Inhibitors of c-Jun N-terminal kinases (JNK) and other protein kinases

ABSTRACT

The present invention provides compounds of formula I:                    
     or a pharmaceutically acceptable derivative o thereof, wherein A, B, R a , R 1 , R 2 , R 3 , and R 4  are as described in the specification. These compounds are inhibitors of protein kinase, particularly inhibitors of JNK, a mammalian protein kinase involved cell proliferation, cell death and response to extracellular stimuli; Lck and Src kinase. The invention also relates to methods for producing these inhibitors. The invention also provides pharmaceutical compositions comprising the inhibitors of the invention and methods of utilizing those compositions in the treatment and prevention of various disorders.

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims benefit of priority to U.S. provisionalapplications 60/283,621 filed Apr. 13, 2001, 60/329,440 filed Oct. 15,2001 and 60/292,974 filed May 23, 2001.

TECHNICAL FIELD OF INVENTION

The present invention relates to inhibitors of protein kinase,especially c-Jun N-terminal kinases (JNK) and the Src-family of kinases,including Lck, which are members of the mitogen-activated protein (MAP)kinase family. JNK, Src, and Lck have been implicated in a number ofdifferent human diseases. The invention also provides pharmaceuticalcompositions comprising the inhibitors of the invention and methods ofutilizing those compositions in the treatment and prevention of variousdisorders in which JNK, Src, and Lck play a role.

BACKGROUND OF THE INVENTION

Mammalian cells respond to extracellular stimuli by activating signalingcascades that are mediated by members of the mitogen-activated protein(MAP) kinase family, which include the extracellular signal regulatedkinases (ERKs), the p38 MAP kinases and the c-Jun N-terminal kinases(JNKs). MAP kinases (MAPKs) are activated by a variety of signalsincluding growth factors, cytokines, UV radiation, and stress-inducingagents. MAPKs are serine/threonine kinases and their activation occur bydual phosphorylation of threonine and tyrosine at the Thr-X-Tyr segmentin the activation loop. MAPKs phosphorylate various substrates includingtranscription factors, which in turn regulate the expression of specificsets of genes and thus mediate a specific response to the stimulus.

One particularly interesting kinase family are the c-Jun NH₂-terminalprotein kinases, also known as JNKs. Three distinct genes, JNK1, JNK2,JNK3 have been identified and at least ten different splicing isoformsof JNKs exist in mammalian cells [Gupta et al., EMBO J., 15:2760-70(1996)]. Members of the JNK family are activated by proinflammatorycytokines, such as tumor necrosis factor-α (TNFα) and interleukin-1 β(IL-1β), as well as by environmental stress, including anisomycin, UWirradiation, hypoxia, and osmotic shock [Minden et al., Biochemica etBiophysica Acta, 1333:F85-F104 (1997)].

The down-Stream substrates of JNKs include transcription factors c-Jun,ATF-2, Elk1, p53 and a cell death domain protein (DENN) [Zhang et al.Proc. Natl. Acad. Sci. USA, 95:2586-91 (1998)]. Each JNK isoform bindsto these substrates with different affinities, suggesting a regulationof signaling pathways by substrate specificity of different JNKs in vivo(Gupta et al., supra).

JNKs, along with other MAPKs, have been implicated in having a role inmediating cellular response to cancer, thrombin-induced plateletaggregation, immunodeficiency disorders, autoimmune diseases, celldeath, allergies, osteoporosis and heart disease. The therapeutictargets related to activation of the JNK pathway include chronicmyelogenous leukemia (CML), rheumatoid arthritis, asthma,osteoarthritis, ischemia, cancer and neurodegenerative diseases.

Several reports have detailed the importance of JNK activationassociated with liver disease or episodes of hepatic ischemia [Nat.Genet. 21:326-9 (1999); FEBS Lett. 420:201-4 (1997); J. Clin. Invest.102:1942-50 (1998); Hepatology 28:1022-30 (1998)]. Therefore, inhibitorsof JNK may be useful to treat various hepatic disorders.

A role for JNK in cardiovascular disease such as myocardial infarctionor congestive heart failure has also been reported as it has been shownJNK mediates hypertrophic responses to various forms of cardiac stress[Circ. Res. 83:167-78 (1998); Circulation 97:1731-7 (1998); J. Biol.Chem. 272:28050-6 (1997); Circ. Res. 79:162-73 (1996); Circ. Res.78:947-53 (1996); J. Clin. Invest. 97:508-14 (1996)].

It has been demonstrated that the JNK cascade also plays a role inT-Cell activation, including activation of the IL-2 promoter. Thus,inhibitors of JNK may have therapeutic value in altering pathologicimmune responses [J. Immunol. 162:3176-87 (1999); Eur. J. Immunol.28:3867-77 (1998); J. Exp. Med. 186:941-53 (1997); Eur. J. Immunol.26:989-94 (1996)].

A role for JNK activation in various cancers has also been established,suggesting the potential use of JNK inhibitors in cancer. For example,constitutively activated JNK is associated with HTLV-1 mediatedtumorigenesis [Oncogene 13:135-42 (1996)]. JNK may play a role inKaposi's sarcoma (KS) because it is thought that the proliferativeeffects of bFGF and OSM on KS cells are mediated by their activation ofthe JNK signaling pathway [J. Clin. Invest. 99:1798-804 (1997)]. Otherproliferative effects of other cytokines implicated in KS proliferation,such as vascular endothelial growth factor (VEGF), IL-6 and TNFα, mayalso be mediated by JNK. In addition, regulation of the c-jun gene inp210 BCR-ABL transformed cells corresponds with activity of JNK,suggesting a role for JNK inhibitors in the treatment for chronicmyelogenous leukemia (CML) [Blood 92:2450-60 (1998)].

JNK1 and JNK2 are widely expressed in a variety of tissues. In contrast,JNK3, is selectively expressed in the brain and to a lesser extent inthe heart and testis [Gupta et al., supra; Mohit et al., Neuron 14:67-78(1995); Martin et al., Brain Res. Mol. Brain Res. 35:47-57 (1996)]. JNK3has been linked to neuronal apoptosis induced by kainic acid, indicatinga role of JNK in the pathogenesis of glutamate neurotoxicity. In theadult human brain, JNK3 expression is localized to a subpopulation ofpyramidal neurons in the CA1, CA4 and subiculum regions of thehippocampus and layers 3 and 5 of the neocortex [Mohit et al., supra].The CA1 neurons of patients with acute hypoxia showed strong nuclearJNK3-immunoreactivity compared to minimal, diffuse cytoplasmic stainingof the hippocampal neurons from brain tissues of normal patients [Zhanget al., supra]. Thus, JNK3 appears to be involved involved in hypoxicand ischemic damage of CA1 neurons in the hippocampus.

In addition, JNK3 co-localizes immunochemically with neurons vulnerablein Alzheimer's disease [Mohit et al., supra]. Disruption of the LNK3gene caused resistance of mice to the excitotoxic glutamate receptoragonist kainic acid, including the effects on seizure activity, AP-1transcriptional activity and apoptosis of hippocampal neurons,indicating that the JNK3 signaling pathway is a critical component inthe pathogenesis of glutamate neurotoxicity (Yang et al., Nature,389:865-870 (1997)].

Based on these findings, JNK signalling, especially that of JNK3, hasbeen implicated in the areas of apoptosis-driven neurodegenerativediseases such as Alzheimer's Disease, Parkinson's Disease, ALS(Amyotrophic Lateral Sclerosis), epilepsy and seizures, Huntington'sDisease, traumatic brain injuries, as well as ischemic and hemorrhagingstroke.

The Src-family of kinases are implicated in cancer, immune systemdysfunction, and bone remodeling diseases. For general reviews, seeThomas and Brugge, Annu. Rev. Cell Dev. Biol. (1997) 13, 513; Lawrenceand Niu, Pharmacol. Ther. (1998) 77, 81; Tatosyan and Mizenina,Biochemistry (Moscow) (2000) 65, 49; Boschelli et al., Drugs of theFuture 2000, 25(7), 717, (2000).

Members of the Src family include the following eight kinases inmammals: Src, Fyn, Yes, Fgr, Lyn, Hck, Lck, Blk and Yrc. These arenonreceptor protein kinases that range in molecular mass from 52 to 62kD. All are characterized by a common structural organization that iscomprised of six distinct functional domains: Src homology domain 4(SH4), a unique domain, SH3 domain, SH2 domain, a catalytic domain(SH1), and a C-terminal regulatory region. Tatosyan et al. Biochemistry(Moscow) 65, 49-58 (2000).

Based on published studies, Src kinases are considered as potentialtherapeutic targets for various human diseases. Mice that are deficientin Src develop osteopetrosis, or bone build-up, because of depressedbone resorption by osteoclasts. This suggests that osteoporosisresulting from abnormally high bone resorption can be treated byinhibiting Src. Soriano et al., Cell, 69, 551 (1992) and Soriano et al.,Cell, 64, 693 (1991).

Suppression of arthritic bone destruction has been achieved by theoverexpression of CSK in rheumatoid synoviocytes and osteoclasts.Takayanagi et al., J. Clin. Invest., 104, 137 (1999). CSK, or C-terminalSrc kinase, phosphorylates and thereby inhibits Src catalytic activity.This implies that Src inhibition may prevent joint destruction that ischaracteristic in patients suffering from rheumatoid arthritis.Boschelli et al., Drugs of the Future 2000, 25(7), 717, (2000).

Src also plays a role in the replication of hepatitis B virus. Thevirally encoded transcription factor HBx activates Src in a steprequired for propagation of the virus. Klein et al., EMBO J., 18, 5019,(1999) and Klein et al., Mol. Cell. Biol., 17, 6427 (1997).

A number of studies have linked Src expression to cancers such as colon,breast, hepatic and pancreatic cancer, certain B-Cell leukemias andlymphomas. Talamonti et al., J. Clin. Invest., 91, 53 (1993); Lutz etal., Biochem. Biophys. Res. 243, 503 (1998); Rosen et al., J. Biol.Chem., 261, 13754 (1986); Bolen et al., Proc. Natl. Acad. Sci. USA, 84,2251 (1987); Masaki et al., Hepatology, 27, 1257 (1998); Biscardi etal., Adv. Cancer Res., 76, 61 (1999); Lynch et al., Leukemia, 7, 1416(1993). Furthermore, antisense Src expressed in ovarian and colon tumorcells has been shown to inhibit tumor growth. Wiener et al., Clin.Cancer Res., 5, 2164 (1999); Staley et al., Cell Growth Diff., 8, 269(1997).

Other Src family kinases are also potential therapeutic targets. Lckplays a role in T-Cell signaling. Mice that lack the Lck gene have apoor ability to develop thymocytes. The function of Lck as a positiveactivator of T-Cell signaling suggests that Lck inhibitors may be usefulfor treating autoimmune disease such as rheumatoid arthritis. Molina etal., Nature, 357, 161 (1992). Hck, Fgr and Lyn have been identified asimportant mediators of integrin signaling in myeloid leukocytes. Lowellet al., J. Leukoc. Biol., 65, 313 (1999). Inhibition of these kinasemediators may therefore be useful for treating inflammation. Boschelliet al., Drugs of the Future 2000, 25(7), 717, (2000).

Accordingly, there is still a great need to develop potent inhibitors ofJNKs and Src family kinases that are useful in treating variousconditions associated with JNK and Src activation.

SUMMARY OF THE INVENTION

The present invention provides compounds of formula I:

wherein R¹, R², R³, R⁴, and R^(a) are as described below.

The present invention also provides a pharmaceutical compositioncomprising a compound of formula I.

The compounds and pharmaceutical compositions of the present inventionare useful as inhibitors of c-Jun N-terminal kinases (JNK) and Srcfamily kinases, including Src and Lck. Thus, they are also useful inmethods for treating or preventing a variety of disorders, such as heartdisease, immunodeficiency disorders, inflammatory diseases, allergicdiseases, autoimmune diseases, destructive bone disorders such asosteoporosis, proliferative disorders, infectious diseases and viraldiseases. The compositions are also useful in methods for preventingcell death and hyperplasia and therefore may be used to treat or preventreperfusion/ischemia in stroke, heart attacks, and organ hypoxia. Thecompositions are also useful in methods for preventing thrombin-inducedplatelet aggregation. The compositions are especially useful fordisorders such as chronic myelogenous leukemia (CML), rheumatoidarthritis, asthma, osteoarthritis, ischemia, cancer, liver diseaseincluding hepatic ischemia, heart disease such as myocardial infarctionand congestive heart failure, pathologic immune conditions involving Tcell activation and neurodegenerative disorders.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides a compound of formula I:

or a pharmaceutically acceptable derivative thereof, wherein:

A and B are each independently selected from N or CH;

R¹ and R² are each independently selected from halogen, CN, NO₂, N(R)₂,OR, SR, or (T)_(n)—R⁵;

R³ is selected from a 3-6 membered carbocyclic or heterocyclic ringhaving one to two heteroatoms independently selected from nitrogen,oxygen, or sulfur, phenyl, or a 5-6 membered heteroaryl ring having oneto three heteroatoms independently selected from nitrogen, oxygen, orsulfur, wherein said phenyl or heteroaryl ring is optionally substitutedwith one (T)_(n)—Ar and one to two R⁷;

each n is independently selected from zero or one;

T is a C₁-C₆ alkylidene chain, wherein one methylene unit of T isoptionally replaced by CO, CO₂, COCO, CONR, OCONR, NRNR, NRNRCO, NRCO,NRCO₂, NRCONR, SO₂, NRSO₂, SO₂NR, NRSO₂NR, O, S, or NR;

each R is independently selected from hydrogen or an optionallysubstituted C₁-C₆ aliphatic group; or two R on the same nitrogen atommay be taken together with the nitrogen to form a four to eightmembered, saturated or unsaturated heterocyclic ring containing one tothree heteroatoms independently selected from nitrogen, oxygen, orsulfur;

R⁴ is (T)_(n)—R, (T)_(n)—Ar, or (T)_(n)—Ar¹;

R^(a) is selected from R^(b), halogen, NO₂, OR^(b), SR^(b), orN(R^(b))₂;

R^(b) is selected from hydrogen or a C₁-C₄ aliphatic group optionallysubstituted with oxo, OH, SH, NH₂, halogen, NO₂, or CN;

R⁵ is an optionally substituted C₁-C₆ aliphatic or Ar;

Ar is a 5-6 membered saturated, partially unsaturated, or arylmonocyclic ring having zero to three heteroatoms independently selectedfrom nitrogen, sulfur, or oxygen, or an 8-10 membered saturated,partially unsaturated, or aryl bicyclic ring having zero to fourheteroatoms independently selected from nitrogen, sulfuri or oxygen,wherein Ar is optionally substituted with one to three R⁷;

Ar¹ is a 6-membered aryl ring having zero to two nitrogens, wherein saidring is substituted with one Z—R⁶ group and optionally substituted withone to three R⁷;

Z is a C₁-C₆ alkylidene chain wherein up to two non-adjacent methyleneunits of Z are optionally replaced by CO, CO₂, COCO, CONR, OCONR, NRNR,NRNRCO, NRCO, NRCO₂, NRCONR, SO, SO₂, NRSO₂, SO₂NR, NRSO₂NR, O, S, orNR; provided that said optionally replaced methylene unit of Z is amethylene unit non-adjacent to R⁶;

R⁶ is selected from Ar, R, halogen, NO₂, CN, OR, SR, N(R)₂, NRC(O)R,NRC(O)N(R)₂, NRCO₂R, C(O)R, CO₂R, OC(O)R, C(O)N(R)₂, OC(O)N(R)₂, SOR,SO₂R, SO₂N(R)₂, NRSO₂R, NRSO₂N(R)₂, C(O)C(O)R, or C(O)CH₂C(O)R; and

each R⁷ is independently selected from R, halogen, NO₂, CN, OR, SR,N(R)₂, NRC(O)R, NRC(O)N(R)₂, NRCO₂R, C(O)R, CO₂R, C(O)N(R)₂, OC(O)N(R)₂,SOR, SO₂R, SO₂N(R)₂, NRSO₂R, NRSO₂N(R)₂, C(O)C(O)R, or C(O)CH₂C(O)R; ortwo R⁷ on adjacent positions of Ar¹ may be taken together to form asaturated, partially unsaturated, or fully unsaturated five to sevenmembered ring containing zero to three heteroatoms selected from O, S,or N.

The following abbreviations are used throughout the specifications(including in chemical formulae):

iPr=isopropyl

t-Bu or tBu=tert-butyl

Et=ethyl

Me=methyl

Cbz=benzoyloxycarbonyl

BOC=tert-butyloxycarbonyl

Ph=phenyl

Bn=benzyl

DMF=N,N-dimethylformamide

THF=tetrahydrofuran

DCM=dichloromethane

DMF-DMA=N,N-dimethylformamide-dimethylacetal

DMSO-dimethylsulfoxide

TLC=thin layer chromatography

As used herein, the following definitions shall apply unless otherwiseindicated.

The phrase “optionally substituted” is used interchangeably with thephrase “substituted or unsubstituted” or with the term“(un)substituted.” Unless otherwise indicated, an optionally substitutedgroup may have a substituent at each substitutable position of thegroup, and each substitution is independent of the other.

The term “aliphatic” or “aliphatic group” as used herein means astraight-chain or branched C₁-C₁₂ hydrocarbon chain that is completelysaturated or that contains one or more units of unsaturation, or amonocyclic C₃-C₈ hydrocarbon or bicyclic C₈-C₁₂ hydrocarbon that iscompletely saturated or that contains one or more units of unsaturation,but which is not aromatic (also referred to herein as “carbocycle” or“cycloalkyl”), that has a single point of attachment to the rest of themolecule wherein any individual ring in said bicyclic ring system has3-7 members. For example, suitable aliphatic groups include, but are notlimited to, linear or branched or alkyl, alkenyl, alkynyl groups andhybrids thereof such as (cycloalkyl)alkyl, (cycloalkenyl)alkyl or(cycloalkyl)alkenyl.

The terms “alkyl”, “alkoxy”, “hydroxyalkyl”, “alkoxyalkyl”, and“alkoxycarbonyl”, used alone or as part of a larger moiety includes bothstraight and branched chains containing one to twelve carbon atoms. Theterms “alkenyl” and “alkynyl” used alone or as part of a larger moietyshall include both straight and branched chains containing two to twelvecarbon atoms.

The terms “haloalkyl”, “haloalkenyl” and “haloalkoxy” means alkyl,alkenyl or alkoxy, as the case may be, substituted with one or morehalogen atoms. The term “halogen” means F, Cl, Br, or I.

The term “heteroatom” means nitrogen, oxygen, or sulfur and includes anyoxidized form of nitrogen and sulfur, and the quaternized form of anybasic nitrogen. Also the term “nitrogen” includes a substitutablenitrogen of a heterocyclic ring. As an example, in a saturated orpartially unsaturated ring having 0-3 heteroatoms selected from oxygen,sulfur or nitrogen, the nitrogen may be N (as in3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl) or NR⁺ (as inN-Substituted pyrrolidinyl).

The term “aryl” used alone or as part of a larger moiety as in“aralkyl”, “aralkoxy”, or “aryloxyalkyl”, refers to monocyclic, bicyclicand tricyclic ring systems having a total of five to fourteen ringmembers, wherein at least one ring in the system is aromatic and whereineach ring in the system contains 3 to 7 ring members. The term “aryl”may be used interchangeably with the term “aryl ring”. The term “aryl”also refers to heteroaryl ring systems as defined hereinbelow.

The term “heterocycle”, “heterocyclyl”, or “heterocyclic” as used hereinmeans non-aromatic, monocyclic, bicyclic or tricyclic ring systemshaving five to fourteen ring members in which one or more ring membersis a heteroatom, wherein each ring in the system contains 3 to 7 ringmembers.

The term “heteroaryl”, used alone or as part of a larger moiety as in“heteroaralkyl” or “heteroarylalkoxy”, refers to monocyclic, bicyclicand tricyclic ring systems having a total of five to fourteen ringmembers, wherein at least one ring in the system is aromatic, at leastone ring in the system contains one or more heteroatoms, and whereineach ring in the system contains 3 to 7 ring members. The term“heteroaryl” may be used interchangeably with the term “heteroaryl ring”or the term “heteroaromatic”.

An aryl (including aralkyl, aralkoxy, aryloxyalkyl and the like) orheteroaryl (including heteroaralkyl and heteroarylalkoxy and the like)group may contain one or more substituents. Suitable substituents on theunsaturated carbon atom of an aryl, heteroaryl, aralkyl, orheteroaralkyl group are selected from halogen, —R°, —OR°, —SR°,1,2-methylene-dioxy, 1,2-ethylenedioxy, phenyl (Ph) optionallysubstituted with R°, —O(Ph) optionally substituted with R°, —CH₂(Ph)optionally substituted with R°, —CH₂CH₂(Ph), optionally substituted withR°, —NO₂, —CN, —N(R°)₂, —NR°C(O)R°, —NR°C(O)N(R°)₂, —NR°CO₂R°,—NR°NR°C(O)R°, —NR°NR°C(O)N(R°)₂, —NR°NR°CO₂R°, —C(O)C(O)R°,—C(O)CH₂C(O)R°, —CO₂R°, —C(O)R°, —C(O)N(R°)₂, —OC(O)N(R°)₂, —S(O)₂R°,—SO₂N(R°)₂, —S(O)R°, —NR°SO₂N(R°)₂, —NR°SO₂R°, —C(═S)N(R°)₂,—C(═NH)—N(R°)₂, or —(CH₂)_(y)NHC(O)R°, wherein each R° is independentlyselected from hydrogen, optionally substituted C₁₋₆ aliphatic, anunsubstituted 5-6 membered heteroaryl or heterocyclic ring, phenyl,—O(Ph), or —CH₂(Ph). Optional substituents on the aliphatic group of R°are selected from NH₂, NH(C₁₋₄ aliphatic), N(C₁₋₄ aliphatic)₂, halogen,C₁₋₄ aliphatic, OH, O(C₁₋₄ aliphatic), NO₂, CN, CO₂H, CO₂(C₁₋₄aliphatic), O(halo C₁₋₄ aliphatic), or halo C₁₋₄ aliphatic.

An aliphatic group or a non-aromatic heterocyclic ring may contain oneor more substituents. Suitable substituents on the saturated carbon ofan aliphatic group or of a non-aromatic heterocyclic ring are selectedfrom those listed above for the unsaturated carbon of an aryl orheteroaryl group and the following: ═O, ═S, ═NNHR*, ═NN(R*)₂,═NNHC(O)R*, ═NNHCO₂(alkyl), ═NNHSO₂(alkyl), or ═NR*, where each R* isindependently selected from hydrogen or an optionally substituted C₁₋₆aliphatic. Optional substituents on the aliphatic group of R* areselected from NH₂, NH(C₁₋₄ aliphatic), N(C₁₋₄ aliphatic)₂, halogen, C₁₋₄aliphatic, OH, O(C₁₋₄ aliphatic), NO₂, CN, CO₂H, CO₂(C₁₋₄ aliphatic),O(halo C₁₋₄ aliphatic), or halo(C₁₋₄ aliphatic).

Optional substituents on the nitrogen of a non-aromatic heterocyclicring are selected from —R⁺, —N(R⁺)₂, —C(O)R⁺, —CO₂R⁺, —C(O)C(O)R⁺,—C(O)CH₂C(O)R⁺, —SO₂R⁺, —SO₂N(R⁺)₂, —C(═S)N(R⁺)₂, —C(═NH)—N(R⁺)₂, or—NR⁺SO₂R⁺; wherein R⁺ is hydrogen, an optionally substituted C₁₋₆aliphatic, optionally substituted phenyl, optionally substituted —O(Ph),optionally substituted —CH₂(Ph), optionally substituted-CH₂CH₂(Ph), oran unsubstituted 5-6 membered heteroaryl or heterocyclic ring. Optionalsubstituents on the aliphatic group or the phenyl ring of R⁺ areselected from NH₂, NH(C₁₋₄ aliphatic), N(C₁₋₄ aliphatic)₂, halogen, C₁₋₄aliphatic, OH, O(C₁₋₄ aliphatic), NO₂, CN, CO₂H, CO₂(C₁₋₄ aliphatic),O(halo C₁₋₄ aliphatic), or halo(C₁₋₄ aliphatic).

The term “alkylidene chain” refers to a straight or branched carbonchain that may be fully saturated or have one or more units ofunsaturation.

A combination of substituents or variables is permissible only if such acombination results in a stable or chemically feasible compound. Astable compound or chemically feasible compound is one that is notsubstantially altered when kept at a temperature of 40° C. or less, inthe absence of moisture or other chemically reactive conditions, for atleast a week.

It will be apparent to one skilled in the art that certain compounds ofthis invention may exist in tautomeric forms, all such tautomeric formsof the compounds being within the scope of the invention.

Unless otherwise stated, structures depicted herein are also meant toinclude all stereochemical forms of the structure; i.e., the R and Sconfigurations for each asymmetric center. Therefore, singlestereochemical isomers as well as enantiomeric and diastereomericmixtures of the present compounds are within the scope of the invention.Unless otherwise stated, structures depicted herein are also meant toinclude compounds which differ only in the presence of one or moreisotopically enriched atoms. For example, compounds having the presentstructures except for the replacement of a hydrogen by a deuterium ortritium, or the replacement of a carbon by a ¹³C- or ¹⁴C-enriched carbonare within the scope of this invention.

Preferred R¹ groups of formula I are selected from N(R)₂, OR, SR, or(T)_(n)—R⁵ wherein T is a C₁₋₄ alkylidene chain and wherein onemethylene unit of T is optionally replaced by S, O, N(R), or CO₂. Morepreferred RI groups of formula I are selected from SCH₂-4-phenol, SCH₃,OH, OEt, N(Me)₂, OMe, 4-methylpiperidin-1-yl, NHEt,NHCH₂CH₂piperidin-1-yl, or NHCH₂CH₂morpholin-4-yl.

Preferred R² groups of formula I are selected from CN, R⁵, halogen,CO₂R⁵, or N(R)₂. More preferred R² groups are selected from CN or CO₂R⁵.

Preferred R³ groups of formula I are selected from 5-6 membered ringselected from carbocyclic, phenyl, or a heterocyclyl or heteroaryl ringhaving one to two heteroatoms independently selected from nitrogen,oxygen or sulfur, wherein R³ is optionally substituted with one(T)_(n)—Ar group and one R⁷. More preferred R³ groups of formula I areselected from phenyl, pyridyl, pyrimidinyl, cyclohexyl, and furanyl.Preferred substituents on R³ are selected from (T)_(n)—Ar or R⁷ whereinAr is a an optionally substituted 5-6 membered aryl ring having zero totwo heteroatoms independently selected from nitrogen, oxygen, or sulfur,and wherein R⁷ is selected from R, halogen, OR, N(R)₂, or CO₂R. Morepreferred substituents on R³ are selected from phenyl, phenoxy, benzyl,benzyloxy, pyridyl, 3-hydroxyphenyl, 2 -hydroxyphenyl, 3-aminophenyl,N-BOC-pyrrolyl, 4-chlorophenyl, 3-ethoxypyridyl, 2-methoxypyridyl,2,5-dimethylisoxazolyl, 3-ethoxyphenyl, 4-isopropylphenyl,4-F-3-Cl-phenyl, pyrrolyl, pyrimidinyl, halogen, such as chloro, bromo,and fluoro, haloalkyl such as trifluoromethyl, OH, NH₂, alkyl, such asmethyl, and alkoxy, such as methoxy and ethoxy.

Preferred R⁴ groups of formula I are selected from hydrogen or Arwherein Ar is an optionally substituted 6 membered saturated, partiallysaturated, or aryl ring having zero to two heteroatoms independentlyselected from nitrogen, oxygen, or sulfur. More preferred R⁴ groups offormula I are selected from phenyl, benzyl, pyridyl, piperidinyl, andcyclohexyl. Preferred substituents on R⁴ are selected from CO₂R, OR,OAr, halogen, NRSO₂R, SO₂N(R)₂, NRCON(R)₂, NO₂, or N(R)₂. More preferredsubstituents of R⁴ are selected from benzyloxy, phenoxy, SO₂NH₂, OH,NO₂, NH₂, OMe, Br, Cl, CO₂Me, NHSO₂Me, NHSO₂Et, NHCON(Me)₂, NHCON(Et)₂,NHCOpyrrolidin-1-yl, or NHCOmorpholin-4-yl.

Most preferred R⁴ groups of formula I are those wherein R⁴ is Ar¹.Preferred Z—R⁶ groups of the Ar¹ group of formula I are those wherein Zis a C₁₋₄ alkylidene chain wherein one methylene unit of Z is optionallyreplaced by O, NH, NHCO, NHCO₂, NHSO₂, CONH, and wherein R⁶ is selectedfrom N(R)₂, NHCOR, or Ar wherein Ar is a 5-6 membered heterocyclic orheteroaryl ring having one to two heteroatoms independently selectedfrom nitrogen, oxygen, or sulfur. The Ar group of R⁶ is optionallysubstituted with R, OR, N(R)₂, or oxo. More preferred Z—R⁶ groups offormula I are selected from O(CH₂)₃OH, O(CH₂)₃NH(CH₂)₂OH,O(CH₂)₂NH(CH₂)₂OH, O(CH₂)₃N(hydroxyethyl) (methyl),O(CH₂)₃pyrrolidin-1-yl, O(CH₂)₂morpholin-4-yl, O(CH₂)₃N(Me)₂,O(CH₂)₃N(Et)₂, O(CH₂)₃ (4-hydroxyethylpiperazin-1-yl),O(CH₂)₃piperazin-1-yl,O(CH₂)₃(4-hydroxymethylpiperidin-1-yl),O(CH₂)₃(4-hydroxypiperidin-1-yl), NHCO(CH₂)₃N(Me)₂, NHCO(CH₂)₃NCOCH₃,NHCOCH₂pyridin-2-yl, NHCOCH₂(2-aminothiazol-4-yl), NHCOCH₂cyclopropyl,NHCO(CH₂)₂N(Et)₂, NHCO(CH₂)₂(piperazin-2,5-dione-3-yl),NHCOpyrrolidin-1-yl, NHCOmorpholin-4-yl, NHCO₂CH₂tetrahydrofuran-2-yl,NHCO₂tetrahydrofuran-2-yl, NHCO₂tetrahydropyran-4-yl, orNHCO₂CH₂tetrahydropyran-2-yl.

Preferred R^(a) groups of formula I are selected from R^(b), OR^(b),SR^(b), or N (R^(b))₂. More preferred R^(a) groups of formula I areselected from methyl, OH, OMe, or NH₂.

One embodiment of this invention relates to compounds of formula IIa:

or a pharmaceutically acceptable derivative thereof, wherein R¹, R², R³,and R⁴ are as described above.

Preferred R¹ groups of formula IIa are selected from N(R)₂, OR, SR, or(T)_(n)—R⁵ wherein T is a C₁₋₄ alkylidene chain and wherein onemethylene unit of T is optionally replaced by S, O, N(R), or CO₂. Morepreferred R¹ groups of formula IIa are selected from SCH₂-4-phenol,SCH₃, OH, OEt, N(Me)₂, OMe, 4-methylpiperidin-1-yl, NHEt,NHCH₂CH₂piperidin-1-yl, or NHCH₂CH₂morpholin-4-yl.

Preferred R² groups of formula IIa are selected from CN, R⁵, halogen,CO₂R⁵, or N(R)₂. More preferred R² groups are selected from CN or CO₂R⁵.

Preferred R³ groups of formula IIa are selected from 5-6 membered ringselected from carbocyclic, phenyl, or a heterocyclyl or heteroaryl ringhaving one to two heteroatoms independently selected from nitrogen,oxygen or sulfur, wherein R³ is optionally substituted with one(T)_(n)—Ar group and one R⁷. More preferred R³ groups of formula IIa areselected from phenyl, pyridyl, pyrimidinyl, cyclohexyl, and furanyl.Preferred substituents on R³ are selected from (T)_(n)—Ar or R⁷ whereinAr is a an optionally substituted 5-6 membered aryl ring having zero totwo heteroatoms independently selected from nitrogen, oxygen, or sulfur,and wherein R⁷ is selected from R, halogen, OR, N(R)₂, or CO₂R. Morepreferred substituents on R³ are selected from phenyl, phenoxy, benzyl,benzyloxy, pyridyl, 3-hydroxyphenyl, 2-hydroxyphenyl, 3-aminophenyl,N-BOC-pyrrolyl, 4-chlorophenyl, 3-ethoxypyridyl, 2-methoxypyridyl,2,5-dimethylisoxazolyl, 3-ethoxyphenyl, 4-isopropylphenyl,4-F-3-Cl-phenyl, pyrrolyl, pyrimidinyl, halogen, such as chloro, bromo,and fluoro, haloalkyl such as trifluoromethyl, OH, NH₂, alkyl, such asmethyl, and alkoxy, such as methoxy and ethoxy.

Preferred R⁴ groups of formula IIa are selected from hydrogen or Arwherein Ar is an optionally substituted 6 membered saturated, partiallysaturated, or aryl ring having zero to two heteroatoms independentlyselected from nitrogen, oxygen, or sulfur. More preferred R⁴ groups offormula IIa are selected from phenyl, benzyl, pyridyl, piperidinyl, andcyclohexyl. Preferred substituents on R⁴ are selected from CO₂R, OR,OAr, halogen, NRSO₂R, SO₂N(R)₂, NRCON(R)₂, NO₂, or N(R)₂. More preferredsubstituents of R⁴ are selected from benzyloxy, phenoxy, SO₂NH₂, OH,NO₂, NH₂, OMe, Br, Cl, CO₂Me, NHSO₂Me, NHSO₂Et, NHCON (Me)₂, NHCON(Et)₂, NHCOpyrrolidin-1-yl, or NHCOmorpholin-4-yl.

Preferred compounds of formula IIa are those having one or more, morepreferably more than one, and most preferably all, of the featuresselected from the group consisting of:

(a) R¹ is selected from N(R)₂, OR, SR, or (T)_(n)—R⁵;

(b) T is a C₁₋₄ alkylidene chain, wherein one methylene unit of T isoptionally replaced by S, O, N(R), or CO₂;

(c) R² is CN, R, halogen, C₂R⁵, or N(R)₂;

(d) R³ is a 5-6 membered ring selected from carbocyclic, phenyl, or aheterocyclyl or heteroaryl ring having one to two heteroatomsindependently selected from nitrogen, oxygen or sulfur, wherein R³ isoptionally substituted with one (T)_(n)—Ar group and one R⁷; and

(e) R⁴ is hydrogen or Ar, wherein Ar is an optionally substituted 6membered saturated, partially saturated, or aryl ring having zero to twoheteroatoms independently selected from nitrogen, oxygen, or sulfur.

More preferred compounds of formula IIa are those having one or more,more preferably more than one, and most preferably all, of the featuresselected from the group consisting of:

(a) R¹ is selected from SCH₂-4-phenol, SCH₃, OH, OEt, N(Me)₂, OMe,4-methylpiperidin-1-yl, NHEt, NHCH₂CH₂piperidin-1-yl, orNHCH₂CH₂morpholin-4-yl;

(b) R² is CN or C₂R⁵;

(c) R³ is selected from phenyl, pyridyl, pyrimidinyl, cyclohexyl, orfuranyl, wherein R³ is optionally substituted with phenyl, phenoxy,benzyl, benzyloxy, pyridyl, 3-hydroxyphenyl, 2-hydroxyphenyl,3-aminophenyl, N-BOC-pyrrolyl, 4-chlorophenyl, 3-ethoxypyridyl,2-methoxypyridyl, 2,5-dimethylisoxazolyl, 3-ethoxyphenyl,4-isopropylphenyl, 4-F-3-Cl-phenyl, pyrrolyl, pyrimidinyl, chloro,bromo, fluoro, trifluoromethyl, OH, NH₂, methyl, methoxy or ethoxy; and

(d) R⁴ is selected from hydrogen or a phenyl, benzyl, pyridyl,piperidinyl, or cyclohexyl ring, wherein said ring is optionallysubsituted with benzyloxy, phenoxy, SO₂NH₂, OH, NO₂, NH₂, OMe, Br, Cl,CO₂Me, NHSO₂Me, NHSO₂Et, NHCON(Me)₂, NHCON(Et)₂, NHCOpyrrolidin-1-yl, orNHCOmorpholin-4-yl.

Another embodiment relates to compounds of formula IIb:

or a pharmaceutically acceptable derivative thereof, wherein R¹, R², R³,and R⁴ are as defined above.

Preferred R¹, R³, and R⁴ groups of formula IIb are those described abovefor compounds of formula IIa.

Preferred R² groups of formula IIb are CN, R⁷, Ar, halogen, or N(R⁶)₂.When R² is Ar, a preferred Ar group is 4-(C₁₋₃ alkyl)-thiazol-2-yl.

Preferred compounds of formula IIb are those having one or more, morepreferably more than one, and most preferably all, of the featuresselected from the group consisting of:

(a) R¹ is selected from N(R)₂, OR, SR, or (T)_(n)—R⁵;

(b) T is a C₁₋₄ alkylidene chain, wherein one methylene unit of T isoptionally replaced by S, O, N(R), or CO₂;

(c) R² is CN, R⁷, Ar, halogen, or N(R⁶)₂;

(d) R³ is a 5-6 membered ring selected from carbocyclic, phenyl, or aheterocyclyl or heteroaryl ring having one to two heteroatomsindependently selected from nitrogen, oxygen or sulfur, wherein R³ isoptionally substituted with one (T)_(n)—Ar group and one R⁷; and

(e) R⁴ is hydrogen or Ar, wherein Ar is an optionally substituted 6membered saturated, partially saturated, or aryl ring having zero to twoheteroatoms independently selected from nitrogen, oxygen, or sulfur.

More preferred compounds of formula IIb are those having one or more,more preferably more than one, 2and most preferably all, of the featuresselected from the group consisting of:

(a) R¹ is selected from SCH₂-4-phenol, SCH₃, OH, OEt, N(Me)₂, OMe,4-methylpiperidin-1-yl, NHEt, NHCH₂CH₂piperidin-1-yl, orNHCH₂CH₂morpholin-4-yl;

(b) R² is CN or 4-(C₁₋₃ alkyl)-thiazol-2-yl;

(c) R³ is selected from phenyl, pyridyl, pyrimidinyl, cyclohexyl, orfuranyl, wherein R³ is optionally substituted with phenyl, phenoxy,benzyl, benzyloxy, pyridyl, 3-hydroxyphenyl, 2-hydroxyphenyl,3-aminophenyl, N-BOC-pyrrolyl, 4-chlorophenyl, 3-ethoxypyridyl,2-methoxypyridyl, 2,5-dimethylisoxazolyl, 3-ethoxyphenyl,4-isopropylphenyl, 4-F-3-Cl-phenyl, pyrrolyl, pyrimidinyl, chloro,bromo, fluoro, trifluoromethyl, OH, NH₂, methyl, methoxy or ethoxy; and

(d) R⁴ is selected from hydrogen or a phenyl, benzyl, pyridyl,piperidinyl, or cyclohexyl ring, wherein said ring is optionallysubsituted with benzyloxy, phenoxy, SO₂NH₂, OH, NO₂, NH₂, OMe, Br, Cl,CO₂Me, NHSO₂Me, NHSO₂Et, NHCON(Me)₂, NHCON(Et)₂, NHCOpyrrolidin-1-yl, orNHCOmorpholin-4-yl.

Exemplary structures of formula IIa are set forth in Table 1 below.

TABLE 1 Compounds of Formula IIa No. IIa- R¹ R² R³ R⁴ 1 SMe CN4-CO₂H-phenyl H 2 SMe CN 4-Cl-phenyl H 3 SMe CN 4-CF₃-phenyl H 4 SMe CN4-CH₃-phenyl H 5 SMe CN 2-Cl-phenyl H 6 SMe CN 4-OCH₃-phenyl H 7 NHCH₂PhCN 4-CF₃-phenyl H 8 NHCH₂Ph CN 2-Cl-phenyl H 9 NHCH₂Ph CN 4-OCH₃-phenylH 10 SMe CN 4-Cl-phenyl Ph 11 OEt CN 4-Cl-phenyl Ph 12 SMe CN4-CF₃-phenyl Ph 13 OEt CN 4-CF₃-phenyl Ph 14 SMe CN 4-CH₃-phenyl Ph 15OEt CN 4-CH₃-phenyl Ph 16 SMe CN H R 17 CH₂CH₂OH CN OPh Et 18 CONHEt CF₃pyridin-3-yl CH₂Ph 19 SCH₂Ph NHEt CONHCH₂Ph COPh 20 CH₂NO₂ CONHEtNH(4-Cl-phenyl) H 21 NHCONH₂ OMe CH₂Ph SO₂Me 22 Et CN thiazol-2-yl Ph 23SMe CN piperidin-1-yl cyclohexyl 24 OCH₂Ph Cl 4-CONHMe-phenyl cyclohexyl25 NHMe NO₂ NHPh H 26 SMe NO₂ NH₂ H 27 OEt NO₂ NHCH₂Ph H 28 NHMe NH₂NHPh Ph 29 SMe NH₂ NH₂ Me 30 OEt NH₂ NHCH₂Ph Me 31 NHMe NHCOEt Ph Ph 32SMe NHCOEt CH₂Ph CH₂Ph 33 OEt NHCOEt CH₂Ph H 34 NHMe CONHMe Ph H 35 SMeCONHMe Ph Ph 36 OEt CONRMe 4-Cl-Ph Ph 37 SMe CN Ph 3-OBn-Ph 38 SMe CN Ph3-SO₂NH₂-Ph 39 S-CH₂- CN Ph Ph phen-4-ol 40 SMe CN Ph 3-OH-Ph 41 SMe CNPh 4-OBn-Ph 42 OH CN Ph 3-OBn-Ph 43 SMe CN cyclohexyl 3,5-OMe-Ph 44 SMeCN cyclohexyl 3-SO₂NH₂-Ph 45 SMe CN cyclohexyl 3-OBn-Ph 46 SMe CNcyclohexyl Ph 47 SMe CN cyclohexyl 4-CO₂Et-Ph 48 SMe CN cyclohexyl3-OH-Ph 49 SMe CN 3-OMe-Ph 3-NO₂-Ph 50 SMe CN 3-OMe-Ph 3-NH₂-Ph 51 SMeCN 3-OH-Ph 3-NO₂-Ph 52 SMe CN 3-OBn-Ph Ph 53 SMe CN 3-OBn-Ph 3-NO₂-Ph 54N(Me)₂ CN 3-OBn-Ph Ph 55 N(Me)₂ CN 3-OBn-Ph 3-NO₂-Ph 56 SMe CN 3-pyridyl3-OBn-Ph 57 SMe CN 3-pyridyl 3-OH-Ph 58 OEt CN Ph Ph 59 SMe CN 3-Br-Ph3-NH₂-Ph 60 N(Me)₂ CN 3-OPh-Ph 3-NH₂-Ph 61 SMe CN 3-OPh-Ph 3-NH₂-Ph 62SMe CN 5-Br-3-pyridyl 3-OBn-Ph 63 4-Me- CN 3-OPh-Ph Ph piperidin- 1-yl64 OH CN 4-tolyl Ph 65 SMe CN 3-OBn-Ph 3-OH-Ph 66 SMe CN 3-OPh-Ph3-OH-Ph 67 SMe CN 3-OH-Ph 3-OH-Ph 68 SMe CN 3-Br-Ph 3-OH-Ph 69 SMe CN3-Br-Ph 3-OBn-Ph 70 NHEt CN Ph 3-OH-Ph 71 SMe CN 3-(3-OH-Ph)-Ph 3-OH-Ph72 SMe CN 3-(3-OEt-Ph)-Ph 3-OH-Ph 73 SMe CN 3-(3-pyridyl)-Ph 3-OH-Ph 74SMe CN 5-Ph-pyridin-3-yl 3-OBn-Ph 75 N(Me)₂ CN 5-Br-3-pyridyl 3-OBn-Ph76 N(Me)₂ CN 5-Ph-3-pyridyl 3-OBn-Ph 77 SMe CO₂Et Ph 3-OR-Ph 78 SMeCO₂Et Ph Ph 79 NHEt CN Ph 4-OH-Ph 80 N(Me)₂ CN 5-Ph-pyridin-3-yl 3-OH-Ph81 SMe CN 3-(3-NH₂-Ph)-Ph 3-OH-Ph 82 SMe CN 3-(3-Cl,4-F-Ph)-Ph 3-OH-Ph83 SMe CN 3-(4-iPr-Ph)-Ph 3-OH-Ph 84 SMe CN 5-Ph-pyridin-3-yl 3-OH-Ph 85SMe CN 5-Ph-pyridin-3-yl 3-NO₂-Ph 86 SMe CN 3-(3-N-Boc- 3-OH-Phpyrrol-2- yl)-Ph 87 SMe CN 3-(4-iPr-Ph)-Ph 3-OH-Ph 88 SMe CN 3-pyridyl3-NHSO₂Me- Ph 89 SMe CN 3-pyridyl 3-NHSO₂Et- Ph 90 NHEt CO₂Et Ph3-OMe-Ph 91 SMe CN 3-pyridyl 3-SO₂NH₂-Ph 92 SMe CN 3-(2-OH-Ph)-Ph3-OH-Ph 93 SMe CN 3-(3-pyrrol-2-yl)-Ph 3-OH-Ph 94 SMe CN3-(6-OMe-pyridin- 3-OH-Ph 2-yl)-Ph 95 SMe CN 3-(5-OMe-pyridin- 3-OH-Ph2-yl)-Ph 96 SMe CN 3-(2,5-Me₂- 3-OH-Ph isoxazol-4-yl)-Ph 97 NH(CH₃)₂ CNPh 3-OH-Ph morpholin- 4-yl 98 NH(CH₃)₂ CN Ph 3-SO₂NH₂-Ph morpholin- 4-yl99 NH(CH₃)₂ CN Ph 4-OH-Ph morpholin- 4-yl 100 NH(CH₃)₃ CN Ph 3-OH-PhN(Et)₂ 101 NH(CH₃)₃ CN Ph 4-OH-Ph N(Et)₂ 102 NH(CH₃)₂ CN Ph 3-OH-Phpiperidin- 1-yl 103 NH(CH₃)₂ CN Ph 3-SO₂NH₂-Ph piperidin- 1-yl 104NH(CH₃)₂ CN Ph 4-OH-Ph piperidin- 1-yl 105 SMe CN 3-(pyridin-4-yl)-Ph3-OH-Ph

Exemplary structures of formula IIb are set forth in Table 2 below.

TABLE 2 Compounds of Formula IIb No. IIb- R¹ R² R³ R⁴ 1 SMe4-CH₃-thiazol-2-yl Me Ph 2 SMe 4-CH₃-thiazol-2-yl Me 4-F-Ph 3 SMe4-CH₃-thiazol-2-yl Me 6-Cl-pyridin-3-yl 4 SMe 4-CH₃-thiazol-2-yl Me3-Cl-Ph 5 SMe 4-CH₃-thiazol-2-yl Me 3-CH₃O-Ph 6 SMe 4-CH₃-thiazol-2-ylMe 3-BnO-Ph 7 SMe CN Me Ph 8 SMe CN Me 4-F-Ph 9 SMe CN Me6-Cl-pyridin-3-yl 10  SMe CN Me 3-Cl-Ph 11  SMe CN Me 3-CH₃O-Ph 12  SMeCN Me 3-BnO-Ph

The above formulae IIa and IIb compounds are those having a pyrimidinering. Compounds of formula I having a pyridine or triazine ring areotherwise structurally similar to the formulae IIa and IIb compounds andare represented by the following general formulae IIIa, IIIb, IVa, andIVb shown below in Table 3.

TABLE 3 Formulae IIIa, IIIb, IVa, and IVb

The compounds shown above in Table 3 are structurally similar tocompounds of formula IIa and IIb where the pyrimidine ring of formulaIIa is replaced by a pyridine (IIIa and IIIb) or triazine ring (IVa andIVb). Accordingly, preferred R¹, R², R³, and R⁴ groups of the compoundsshown above in Table 3 are as described above for the formula IIacompounds.

Exemplary structures of formulae IIIa and IIIb are set forth in Table 4below.

TABLE 4 Compounds of Formulae IIIa and IIIb No. R¹ R² R³ R⁴ IIIa-1 SMeCN 4-CO₂H-phenyl H IIIa-2 SMe CN 4-Cl-phenyl H IIIa-3 SMe CN4-CF₃-phenyl H IIIa-4 SMe CN 4-CH₃-phenyl H IIIa-5 SMe CN 2-Cl-phenyl HIIIa-6 SMe CN 4-OCH₃-phenyl H IIIa-7 NHCH₂Ph CN 4-CF₃-phenyl H IIIa-8NHCH₂Ph CN 2-Cl-phenyl H IIIa-9 NHCH₂Ph CN 4-OCH₃-phenyl H IIIa-10 SMeCN 4-Cl-phenyl Ph IIIa-11 OEt CN 4-Cl-phenyl Ph IIIa-12 SMe CN4-CF₃-phenyl Ph IIIa-13 OEt CN 4-CF₃-phenyl Ph IIIa-14 SMe CN4-CH₃-phenyl Ph IIIa-15 OEt CN 4-CH₃-phenyl Ph IIIa-16 CH₂CH₂OH CN OPhEt IIIa-17 CONHEt CF₃ pyridin-3-yl CH₂Ph IIIa-18 SCH₂Ph NHEt CONHCH₂PhCOPh IIIa-19 CH₂NO₂ CONHEt NH(4-Cl-phenyl) H IIIa-20 NHCONH₂ OMe CH₂PhSO₂Me IIIa-21 Et CN thiazol-2-yl Ph IIIa-22 SMe CN piperidin-1-ylcyclohexyl IIIa-23 OCH₂Ph Cl 4-CONHMe-phenyl cyclohexyl IIIb-1 SMe CNCH₃ Ph IIIb-2 SMe CN CH₃ 4-F-Ph IIIb-3 SMe CN CH₃ 3-CH₃O-Ph IIIb-4 OEtCN 4-CH₃-phenyl Ph IIIb-5 CH₂CH₂OH CN OPh Et IIIb-6 CONHEt CF₃pyridin-3-yl CH₂Ph IIIb-7 SCH₂Ph NHEt CONHCH₂Ph COPh IIIb-8 CH₂NO₂CONHEt NH(4-Cl-phenyl) H IIIb-9 NHCONH₂ OMe CH₂Ph SO₂Me

Exemplary structures of formulae IVa and IVb are set forth in Table 5below.

TABLE 5 Compounds of Formula IVa and IVb No.IV- R¹ R² R³ R⁴ IVa-1 SMe CN4-CO₂H-phenyl H IVa-2 SMe CN 4-Cl-phenyl H IVa-3 SMe CN 4-CF₃-phenyl HIVa-4 SMe CN 4-CH₃-phenyl H IVa-5 SMe CN 2-Cl-phenyl H IVa-6 SMe CN4-OCH₃-phenyl H IVa-7 NHCH₂Ph CN 4-CF₃-phenyl H IVa-8 NHCH₂Ph CN2-Cl-phenyl H IVa-9 NHCH₂Ph CN 4-OCH₃-phenyl H IVa-10 SMe CN 4-Cl-phenylPh IVa-11 OEt CN 4-Cl-phenyl Ph IVa-12 SMe CN 4-CF₃-phenyl Ph IVa-13 OEtCN 4-CF₃-phenyl Ph IVa-14 SMe CN 4-CH₃-phenyl Ph IVa-15 OEt CN4-CH₃-phenyl Ph IVa-16 CH₂CH₂OH CN OPh Et IVa-17 CONHEt CF₃ pyridin-3-ylCH₂Ph IVa-18 SCH₂Ph NHEt CONHCH₂Ph COPh IVa-19 CH₂NO₂ CONHEtNH(4-Cl-phenyl) H IVa-20 NHCONH₂ OMe CH₂Ph SO₂Me IVa-21 Et CNthiazol-2-yl Ph IVa-22 SMe CN piperidin-1-yl cyclohexyl IVa-23 OCH₂Ph Cl4-CONHMe-phenyl cyclohexyl IVb-1 SMe CN CH₃ pyridin-3-yl IVb-2 SMe CNCH₃ Ph IVb-3 SMe CN CH₂CH₃ H IVb-4 CH₂CH₂OH CN OPh Et IVb-5 CONHEt CF₃pyridin-3-yl CH₂Ph IVb-6 SCH₂Ph NHEt CONHCH₂Ph COPh IVb-7 CH₂NO₂ CONHEtNH(4-Cl-phenyl) H IVb-8 NHCONH₂ OMe CH₂Ph SO₂Me IVb-9 Et CN thiazol-2-ylPh IVb-10 SMe CN piperidin-1-yl cyclohexyl IVb-11 OCH₂Ph Cl4-CONHMe-phenyl cyclohexyl

A preferred embodiment of this invention relates to compounds of formulaV:

or a pharmaceutically acceptable derivative thereof, wherein R^(a), R¹,R², R³, and R⁴ are as defined above.

Preferred compounds of formula V include those having one or more, andmost preferably all, of the following features:

(a) R¹ is selected from N(R)₂, OR, SR, or (T)_(n)—R⁵;

(b) T is a C₁₋₄ alkylidene chain, wherein one methylene unit of T isoptionally replaced by S, O, N(R), or CO₂;

(c) R² is CN, R, halogen, C₂R⁵, or N(R)₂;

(d) R³ is a 5-6 membered ring selected from carbocyclic, phenyl, or aheterocyclyl or heteroaryl ring having one to two heteroatomsindependently selected from nitrogen, oxygen or sulfur, wherein R³ isoptionally substituted with one (T)_(n)—Ar group and one R⁷;

(e) R⁴ is hydrogen or Ar, wherein Ar is an optionally substituted 6membered saturated, partially saturated, or aryl ring having zero to twoheteroatoms independently selected from nitrogen, oxygen, or sulfur; and

(f) R^(a) is selected from R^(b), OR^(b), SR^(b), or N(R^(b))₂.

More preferred compounds of formula V are those having one or more, morepreferably more than one, and most preferably all, of the featuresselected from the group consisting of:

(a) R¹ is selected from SCH₂-4-phenol, SCH₃, OH, OEt, N(Me)₂, OMe,4-methylpiperidin-1-yl, NHEt, NHCH₂CH₂piperidin-1-yl, orNHCH₂CH₂morpholin-4-yl;

(b) R² is CN or C₂R⁵;

(c) R³ is selected from phenyl, pyridyl, pyrimidinyl, cyclohexyl, orfuranyl, wherein R³ is optionally substituted with phenyl, phenoxy,benzyl, benzyloxy, pyridyl, 3-hydroxyphenyl, 2-hydroxyphenyl,3-aminophenyl, N-BOC-pyrrolyl, 4-chlorophenyl, 3-ethoxypyridyl,2-methoxypyridyl, 2,5-dimethylisoxazolyl, 3-ethoxyphenyl,4-isopropylphenyl, 4-F-3-Cl-phenyl, pyrrolyl, pyrimidinyl, chloro,bromo, fluoro, trifluoromethyl, OH, NH₂, methyl, methoxy or ethoxy;

(d) R⁴ is selected from hydrogen or a phenyl, benzyl, pyridyl,piperidinyl, or cyclohexyl ring, wherein said ring is optionallysubsituted with benzyloxy, phenoxy, SO₂NH₂, OH, NO₂, NH₂, OMe, Br, Cl,CO₂Me, NHSO₂Me, NHSO₂Et, NHCON(Me)₂, NHCON(Et)₂, NHCOpyrrolidin-1-yl, orNHCOmorpholin-4-yl; and

(e) R^(a) is methyl, OH, OMe, or NH₂.

Exemplary structures of formula V, wherein R² is CN, are set forth inTable 7 below.

TABLE 7 Compounds of Formula V No.V- R¹ R^(a) R³ R⁴ 1 SCH₂phen-4-ol Me3-Cl phenyl Ph 2 OH Me Ph 3-OBn-Ph 3 OEt Me Ph 3-OBn-Ph 4 SMe Me Ph3-OBn-Ph 5 SMe Me Ph 3-SO₂NH₂-Ph 6 SMe Me Ph 3-OH-Ph 7 SMe Me Ph3-NO₂-Ph 8 SMe Me Ph 3-NH₂-Ph 9 SMe Me 2-CF₃-Ph 3-OBn-Ph 10 SMe Me2-CF₃-Ph 3-OH-Ph 11 SCH₂phen-4-ol Me 3-Cl-Ph Ph 12 SMe Me 4-Me-Ph3-OBn-Ph 13 SMe Me 4-Me-Ph 3-OH-Ph 14 SMe Me 4-Me-Ph pyrid-3-yl 15 SMeMe pyrid-3-yl 3-OBn-Ph 16 SMe Me pyrid-3-yl Ph 17 SMe Me pyrid-3-yl3-OMe-Ph 18 SMe Me pyrid-3-yl 3,5-OMe-Ph 19 SMe Me pyrid-3-yl 3-Br-Ph 20SMe Me pyrid-3-yl 3-Cl-Ph 21 SMe Me pyrid-3-yl 3-CO₂Me-Ph 22 SMe Mepyrid-3-yl 6-Cl-pyrid-3-yl 23 SMe Me pyrid-3-yl CH₂Ph 24 SMe Mepyrid-3-yl 3-OH-Ph 25 SMe Me furan-2-yl pyrid-3-yl 26 SMe Me furan-2-yl3-OH-Ph 27 SMe Me furan-2-yl 3-OBn-Ph 28 SMe Me furan-2-yl 3-NO₂-Ph 29N(Me)₂ Me 3-OPh-Ph 3-NO₂-Ph 30 N(Me)₂ Me 3-OPh-Ph 3-OH-Ph 31 SMe Me3-Ph-Ph 3-OBn-Ph 32 SMe Me 3-Ph-Ph 3-OH-Ph

A more preferred embodiment relates to compounds of formula VI:

or a pharmaceutically acceptable derivative thereof, wherein R¹, R², R³,R^(a), Z, and R⁶ are as defined above.

Preferred R¹, R², R³, and R^(a) groups of formula VI are those describedabove for formula IIa.

Preferred Z—R⁶ groups of formula VI are those wherein Z is a C₁₋₄alkylidene chain wherein one methylene unit of Z is optionally replacedby O, NH, NHCO, NHCO₂, NHSO₂, CONH, and wherein R⁶ is selected fromN(R)₂, NHCOR, or Ar wherein Ar is a 5-6 membered heterocyclic orheteroaryl ring having one to two heteroatoms independently selectedfrom nitrogen, oxygen, or sulfur. The Ar group of R⁶ is optionallysubstituted with R, OR, N(R)₂, or oxo. More preferred Z—R⁶ groups offormula VI are selected from O(CH₂)₃OH, O(CH₂)₃NH(CH₂)₂OH,O(CH₂)₂NH(CH₂)₂OH, O(CH₂)₃N(hydroxyethyl) (methyl),O(CH₂)₃pyrrolidin-1-yl, O(CH₂)₂morpholin-4-yl, O(CH₂)₃N(Me)₂,O(CH₂)₃N(Et)₂, O(CH₂)₃(4-hydroxyethylpiperazin-1-yl),O(CH₂)₃piperazin-1-yl, O(CH₂)₃(4-hydroxymethylpiperidin-1-yl),O(CH₂)₃(4-hydroxypiperidin-1-yl), NHCO(CH₂)₃N(Me)₂, NHCO(CH₂)₃NCOCH₃,NHCOCH₂pyridin-2-yl, NHCOCH₂(2-aminothiazol-4-yl), NHCOCH₂cyclopropyl,NHCO(CH₂)₂N(Et)₂, NHCO(CH₂)₂(piperazin-2,5-dione-3-yl),NHCOpyrrolidin-1-yl, NHCOmorpholin-4-yl, NHCO₂CH₂tetrahydrofuran-2-yl,NHCO₂tetrahydrofuran-2-yl, NHCO₂tetrahydropyran-4-yl, orNHCO₂CH₂tetrahydropyran-2-yl.

Preferred compounds of formula VI are those having one or more, morepreferably more than one, and most preferably all, of the featuresselected from the group consisting of:

(a) R¹ is N(R)₂, OR, SR, or (T)_(n)—R⁵;

(b) T is a C₁₋₄ alkylidene chain, wherein one methylene unit of T isoptionally replaced by S, O, N(R), or CO₂;

(c) R² is CN, R⁷, halogen, or N(R⁶)₂;

(d) R³ is a 5-6 membered ring selected from carbocyclic, phenyl, or aheterocyclyl or heteroaryl ring having one to two heteroatomsindependently selected from nitrogen, oxygen or sulfur, wherein R³ isoptionally substituted with one (T)_(n)—Ar group and one R⁷;

(e) Z is a C₁₋₄ alkylidene chain wherein one methylene unit of Z isoptionally replaced by O, NH, NHCO, NHCO₂, NHSO₂, CONH;

(f) R⁶ is selected from N(R)₂, NHCOR, or Ar wherein Ar is an optionallysubstituted 5-6 membered heterocyclic or heteroaryl ring having one totwo heteroatoms independently selected from nitrogen, oxygen, or sulfur;and

(g) R^(a) is R^(b), OR^(b), SR^(b), or N(R^(b))₂.

More preferred compounds of formula VI are those having one or more,more preferably more than one, and most preferably all, of the featuresselected from the group consisting of:

(a) R¹ is selected from SCH₂-4-phenol, SCH₃, OH, OEt, N(Me)₂, OMe,4-methylpiperidin-1-yl, NHEt, NHCH₂CH₂piperidin-1-yl, orNHCH₂CH₂morpholin-4-yl;

(b) R² is CN;

(c) R³ is a phenyl, pyridyl, furyl, or cyclohexyl ring optionallysubstituted with (T)_(n)—Ar or R⁷ wherein Ar is a 5-6 membered aryl ringhaving zero to two heteroatoms independently selected from nitrogen,oxygen, or sulfur, and wherein R⁷ is selected from R, halogen, OR,N(R)₂, or CO₂R;

(d) R^(a) is hydrogen or methyl; and

(e) Z—R⁶ is selected from O(CH₂)₃OH, O(CH₂)₃NH(CH₂)₂OH,O(CH₂)₂NH(CH₂)₂OH, O(CH₂)₃N(hydroxyethyl) (methyl),O(CH₂)₃pyrrolidin-1-yl, O(CH₂)₂morpholin-4-yl, O(CH₂)₃N(Me)₂,O(CH₂)₃N(Et)₂, O(CH₂)₃(4-hydroxyethylpiperazin-1-yl),O(CH₂)₃piperazin-1-yl, O(CH₂)₃(4-hydroxymethylpiperidin-1-yl),O(CH₂)₃(4-hydroxypiperidin-1-yl), NHCO (CH₂)₃N(Me)₂, NHCO(CH₂)₃NCOCH₃,NHCOCH₂pyridin-2-yl, NHCOCH₂(2-aminothiazol-4-yl), NHCOCH₂cyclopropyl,NHCO(CH₂)₂N (Et)₂, NHCO(CH₂)₂(piperazin-2,5-dione-3-yl),NHCOpyrrolidin-1-yl, NHCOmorpholin-4-yl, NHCO₂CH₂tetrahydrofuran-2-yl,NHCO₂tetrahydrofuran-2-yl, NHCO₂tetrahydropyran-4-yl, orNHCO₂CH₂tetrahydropyran-2-yl.

Exemplary structures of formula VI are set forth in Table 8 below.

TABLE 8 Compounds of Formula VI

The above formula VI compounds are those having a pyrimidine ring.Compounds of formula VI having a pyridine or triazine ring are otherwisestructurally similar to the formula VI compounds and are represented bythe following general formulae VII and VIII shown below:

The compounds of formulae VII and VIII shown above are structurallysimilar to compounds of formula VI where the pyrimidine ring of formulaVI is replaced by a pyridine (VII) or triazine ring (VIII). Accordingly,preferred R¹, R², R³, and Z—R⁶ groups of the compounds of formulae VIIand VIII are as described above for the formula VI compounds.

The present compounds may be prepared in general by methods known tothose skilled in the art for analogous compounds, as illustrated by thegeneral Schemes I, II, III, IV, V, and VI, and the synthetic examplesshown below.

Scheme I above shows a general synthetic route that is used forpreparing the compounds of formula IIa where R² is CN. These compoundsare prepared in a parallel fashion in the following manner. In step (a),α-cyanoacetophenone (1) is combined with K₂CO₃ (3 equivalents) in DMFand the mixture allowed to stir at room temperature. CS₂ (1.5equivalents) is then added and the resulting mixture stirred at roomtemperature for an additional 10 minutes. A solution of1-chloro-propan-2-one (1.0 equivalent) in DMF is added, then a solutionof MeI (1.1 equivalents) in DMF is added in a dropwise fashion. After 30minutes, the mixture is poured onto water and the resulting mixturestirred vigorously for 12-16 hours to afford a suspension of compound 2.The crude product 2 is isolated by filtration. This reaction may be usedto obtain compounds of this invention derived from α-cyanoacetophenoneshaving a wide variety of phenyl substituents. Examples of suitable R³groups include, but are not limited to, those set forth in Table 1above.

In step (b), the crude product 2 is combined witht-butoxybisdimethylaminomethane (Brederick's reagent, 3) in THF andallowed to stir at room temperature for 12-16 hours. The reactionmixture is concentrated then used directly for step (c). The crudeconcentrate 4 is dissolved in EtOH. Compound 5 is added to the ethanolicsolution and the resulting mixture heated to 90° C. for 4 hours.Although Scheme I uses phenyl guanidine at step (c), it would be obviousto one of skill in the art that other aryl guanidines may be used atstep (c) to prepare compounds of the present invention where R⁴ is avariety of optionally substituted aryl groups. The reaction mixture isconcentrated then, after aqueous work-up, the product is purified bypreparatory HPLC to afford compounds 6 and 7. The details of theconditions used for producing these compounds are set forth in theExamples.

Scheme II above shows a general synthetic route that is used forpreparing the compounds of formula IIb where R³ is methyl and R⁴ is anoptionally substituted aryl group.

Intermediate 4 is prepared by treating 2,4-pentanedione (1) withpotassium carbonate (3 equivalents), CS₂ (2) (1.5 equivalents) andchloroacetonitrile (1.0 equivalents) in DMF at room temperature for 2hours. The mixture is cooled to 0° C. then methyl iodide (3) is addedslowly and the resulting mixture stirred at room temperature for 12hours. Water is added to precipitate the product which is isolated byfiltration to afford 4.

Intermediate 5 is prepared by treating 4 with dimethylformamide-dimethylacetal (DMF-DMA) in acetonitrile at reflux for 18 hours. The product 5is isolated as a yellow solid from trituration with ether.

Compounds of formula IIb are prepared from 5 by combining 5 with an arylguanidine in acetonitrile and heating the resulting mixture at refluxfor 24 hours. Methanol is added to the reaction mixture to precipitatethe product and the resulting suspension filterred to afford productIIb. The details of the conditions used for producing these compoundsare set forth in the Examples.

Scheme III above shows a general synthetic route that is used forpreparing the compounds of formula IIb where R³ is methyl, R⁴ is anoptionally substituted aryl group, and R² is 4-methylthiazol-2-yl.

1-[4-Methyl-2-methylsulfanyl-5-(4-methylthiazol-2-yl)-thiophen-3-yl]-ethanone(1) (Maybridge Chemicals) is treated with DMF-DMA in acetonitrile atreflux for 18 hours. The mixture is concentrated in vacuo and theresidue triturated with diethyl ether to afford 2 as a yellow solid.

Intermediate 2 is combined with an aryl guanidine in acetonitrile andthe resulting mixture heated at reflux for 24 hours. After cooling toroom temperature, methanol is added to precipitate the product. Theproduct 3 is isolated by filtration after methanol washes. The detailsof the conditions used for producing these compounds are set forth inthe Examples.

Using the preparation of compound V-1 as an example, Scheme IV aboveshows a general synthetic route that may be used to prepare compounds offormula V in parallel fashion in the following manner. In step (a), CS₂is added to a slurry of 3-chlorobenzoylacetonitrile (1) and LiOH—H₂O inDMF. The resulting mixture is treated with 1-bromopentan-2-one then tothis mixture is then added bromo-Wang resin. The solvent is removed byfiltration and the resin rinsed with solvent and dried under nitrogen toafford resin-bound compound 2.

In step (b), compound 2 is combined with THF and DMF-DMA and theresulting slurry heated at 60° C. for 18 hours. The solvent is removedby filtration and the resin washed with solvent then dried undernitrogen to afford resin-bound compound 3.

The pyrimidine ring is formed in step (c) by treating 3 with N-phenylguanidine in THF at reflux for 24 hours. The solvent is removed byfiltration and the resin washed several times with solvent. Theresulting resin is again treated with N-phenyl guanidine in THF atreflux for another 24 hours. The solvent is again removed by filtrationand the resin washed several times with solvent then dried undernitrogen to afford resin-bound compound 4.

The product V-1 is cleaved from the resin in step (d) by treating 4 withtrifluoroacetic acid in dichloromethane and water for 3 hours at roomtemperature. The resin is filtered and washed with dichloromethane thentreated with trifluoroacetic acid. The resulting mixture is allowed tosit for 14 hours at room temperature then filtered. The resin is washedwith dichloromethane, the solvent concentrated, and the crude productpurified by preparative HPLC to afford compound V-1. The details of theconditions used for producing these compounds are set forth in theExamples.

Scheme V above shows a method for preparing the intermediate compound 5which may be used to prepare compounds of formulae IIa, IIIa, IVa, V,and VI wherein R³ is a cyclohexyl ring. Intermediate compound 5 may bereadily transformed to compounds of formulae IIa, IIIa, IVa, V, and VIby the methods shown in Schemes I-IV above.

In step (a), a solution of ethyl cyanoacetate (2) in acetonitrile istreated with MgCl₂ and Et₃N at 0° C., and the resulting suspensionstirred at 0° C. for 30 minutes. To this suspension is addedcyclohexanecarbonyl chloride (1) and the reaction mixture stirred at 0°C. Aqueous workup affords 3. A solution of compound 3 in DMSO/H₂O isheated then aqueous work-up affords compound 4.

In step (c), compound 4 may be used to prepare thiophene compound 5 byusing the method described in step (a) of Scheme II above.

Using the preparation of compound VI-18 as an example, Scheme VI aboveshows a method for preparing compounds of formula VI where R³ ispyridin-3-yl and Z is a C₁₋₄ alkylidene chain wherein one methylene unitof Z is replaced by oxygen.

In step (a), a solution of ethyl nicotinate in toluene is treated withNaH and the resulting suspension heated at 90° C. while addingacetonitrile. After heating the reaction overnight, the reaction mixtureis allowed to cool and the resulting solids collected by filtration.

Steps (b), (c), and (d) may be performed in a manner substantiallysimilar to those described in step (a) of Scheme II, step (b) of SchemeIV, and step (c) of Schemes I and II. The resulting compound 6 may beused to prepare a variety of compounds of formula VI by using methodsknown in the art. Compound VI-18 was prepared, via steps (e) through(g), from compound 6 by alkylating the phenol with 3-bromopropanol atstep (e). One of skill in the art would recognize that the compound 6phenol group may be readily derivatized in a number of ways to formother compounds of formula VI. In step (f), the propanol —OH is treatedwith MsCl to form the mesylate which is displaced with dimethyl amine atstep (g) to form compound VI-18. One of skill in the art would recognizethat other leaving groups may be utilized to allow for furtherderivatization of the —OH group and that this leaving group may bedisplaced with a variety of amines to form other compounds of formulaVI. The details of the conditions used to prepared the compounds ofScheme VI are set forth in the Examples.

Scheme VII above shows a general method for preparing compounds offormula VI where Z is a C₁₋₄ alkylidene chain wherein one methylene unitof Z is replaced by NH or NHCO. Compound 1 may be prepared according tothe general methods described above. Compound V-7 may be prepared bysteps (a) and (b) according to the methods described above. In step (c),the nitro group is hydrogenated in the presence of palladium on carbonto form the amino compound V-8. Compound V-8 may be used to prepare avariety of compounds of formula VI. For example, compound V-8 may becoupled with a carboxylic acid in order to form amide compounds 5.Alternatively, compound V-8 may be alkylated to form compounds 6. One ofskill in the art would recognize that compound V-8 may be treated with avariety of reagents to form other compounds of formula VI.

According to another embodiment, the invention provides a method ofinhibiting JNK, Src, or Lck kinase activity in a biological sample. Thismethod comprises the step of contacting said biological sample with acompound of formula I. According to a preferred embodiment, theinvention relates to a method of inhibiting JNK, Src, or Lck kinaseactivity in a biological sample comprising the step of contacting saidbiological sample with a compound of formula IIa, IIb, V, or VI. A morepreferred embodiment relates to contacting said biological sample with acompound of formula IIa or VI.

The term “biological sample”, as used herein, includes, withoutlimitation, cell cultures or extracts thereof; biopsied materialobtained from a mammal or extracts thereof; and blood, saliva, urine,feces, semen, tears, or other body fluids or extracts thereof.

Inhibition of JNK, Src, or Lck kinase activity in a biological sample isuseful for a variety of purposes which are known to one of skill in theart. Examples of such purposes include, but are not limited to, bloodtransfusion, organ-transplantation, biological specimen storage, andbiological assays.

Compounds of formula I or salts thereof may be formulated intocompositions. In a preferred embodiment, the composition is apharmaceutically acceptable composition. In one embodiment, thecomposition comprises an amount of compound effective to inhibit aprotein kinase, particularly JNK, Src, or Lck, in a biological sample orin a patient. In another embodiment, compounds of this invention andpharmaceutical compositions thereof, which comprise an amount of thecompound effective to treat or prevent an JNK, Src, or Lck-mediatedcondition and a pharmaceutically acceptable carrier, adjuvant, orvehicle, may be formulated for administration to a patient.

The amount effective to inhibit protein kinase, for example, JNK, Src,or Lck, is one that measurably inhibits the kinase activity wherecompared to the activity of the enzyme in the absence of an inhibitor.Any method may be used to determine inhibition.

The term “pharmaceutically acceptable carrier, adjuvant, or vehicle”refers to a non-toxic carrier, adjuvant, or vehicle that may beadministered to a patient, together with a compound of this invention,and which does not destroy the pharmacological activity thereof.

The term “patient” includes human and veterinary subjects.

Pharmaceutically acceptable carriers that may be used in thesepharmaceutical compositions include, but are not limited to, ionexchangers, alumina, aluminum stearate, lecithin, serum proteins, suchas human serum albumin, buffer substances such as phosphates, glycine,sorbic acid, potassium sorbate, partial glyceride mixtures of saturatedvegetable fatty acids, water, salts or electrolytes, such as protaminesulfate, disodium hydrogen phosphate, potassium hydrogen phosphate,sodium chloride, zinc salts, colloidal silica, magnesium trisilicate,polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol,sodium carboxymethylcellulose, polyacrylates, waxes,polyethylene-polyoxypropylene-block polymers, polyethylene glycol andwool fat.

The compositions of the present invention may be administered orally,parenterally, by inhalation spray, topically, rectally, nasally,buccally, vaginally or via an implanted reservoir. The term “parenteral”as used herein includes subcutaneous, intravenous, intramuscular,intra-articular, intra-synovial, intrasternal, intrathecal,intrahepatic, intralesional and intracranial injection or infusiontechniques. Preferably, the compositions are administered orally,intraperitoneally or intravenously.

Sterile injectable forms of the compositions of this invention may beaqueous or oleaginous suspension. These suspensions may be formulatedaccording to techniques known in the art using suitable dispersing orwetting agents and suspending agents. The sterile injectable preparationmay also be a sterile injectable solution or suspension in a non-toxicparenterally-acceptable diluent or solvent, for example as a solution in1,3-butanediol. Among the acceptable vehicles and solvents that may beemployed are water, Ringer's solution and isotonic sodium chloridesolution. In addition, sterile, fixed oils are conventionally employedas a solvent or suspending medium. For this purpose, any bland fixed oilmay be employed including synthetic mono- or di-glycerides. Fatty acids,such as oleic acid and its glyceride derivatives are useful in thepreparation of injectables, as are natural pharmaceutically-acceptableoils, such as olive oil or castor oil, especially in theirpolyoxyethylated versions. These oil solutions or suspensions may alsocontain a long-chain alcohol diluent or dispersant, such ascarboxymethyl cellulose or similar dispersing agents which are commonlyused in the formulation of pharmaceutically acceptable dosage formsincluding emulsions and suspensions. Other commonly used surfactants,such as Tweens, Spans and other emulsifying agents or bioavailabilityenhancers which are commonly used in the manufacture of pharmaceuticallyacceptable solid, liquid, or other dosage forms may also be used for thepurposes of formulation.

The pharmaceutical compositions of this invention may be orallyadministered in any orally acceptable dosage form including, but notlimited to, capsules, tablets, aqueous suspensions or solutions. In thecase of tablets for oral use, carriers commonly used include lactose andcorn starch. Lubricating agents, such as magnesium stearate, are alsotypically added. For oral administration in a capsule form, usefuldiluents include lactose and dried cornstarch. When aqueous suspensionsare required for oral use, the active ingredient is combined withemulsifying and suspending agents. If desired, certain sweetening,favoring or coloring agents may also be added.

Alternatively, the pharmaceutical compositions of this invention may beadministered in the form of suppositories for rectal administration.These can be prepared by mixing the agent with a suitable non-irritatingexcipient which is solid at room temperature but liquid at rectaltemperature and therefore will melt in the rectum to release the drug.Such materials include cocoa butter, beeswax and polyethylene glycols.

The pharmaceutical compositions of this invention may also beadministered topically, especially when the target of treatment includesareas or organs readily accessible by topical application, includingdiseases of the eye, the skin, or the lower intestinal tract. Suitabletopical formulations are readily prepared for each of these areas ororgans.

Topical application for the lower intestinal tract can be effected in arectal suppository formulation (see above) or in a suitable enemaformulation. Topically-transdermal patches may also be used.

For topical applications, the pharmaceutical compositions may beformulated in a suitable ointment containing the active componentsuspended or dissolved in one or more carriers. Carriers for topicaladministration of the compounds of this invention include, but are notlimited to, mineral oil, liquid petrolatum, white petrolatum, propyleneglycol, polyoxyethylene, polyoxypropylene compound, emulsifying wax andwater. Alternatively, the pharmaceutical compositions can be formulatedin a suitable lotion or cream containing the active components suspendedor dissolved in one or more pharmaceutically acceptable carriers.Suitable carriers include, but are not limited to, mineral oil, sorbitanmonostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol,2-octyldodecanol, benzyl alcohol and water.

For ophthalmic use, the pharmaceutical compositions may be formulated asmicronized suspensions in isotonic, pH adjusted sterile saline, or,preferably, as solutions in isotonic, pH adjusted sterile saline, eitherwith or without a preservative such as benzylalkonium chloride.Alternatively, for ophthalmic uses, the pharmaceutical compositions maybe formulated in an ointment such as petrolatum.

The pharmaceutical compositions of this invention may also beadministered by nasal aerosol or inhalation. Such compositions areprepared according to techniques well-known in the art of pharmaceuticalformulation and may be prepared as solutions in saline, employing benzylalcohol or other suitable preservatives, absorption promoters to enhancebioavailability, fluorocarbons, and/or other conventional solubilizingor dispersing agents.

According to a preferred embodiment, the pharmaceutical compositions ofthis invention are orally administered.

According to another embodiment, the present invention relates to apharmaceutically acceptable derivative of a compound of formula I. In apreferred embodiment, said pharmaceutically acceptable derivative is ofa compound of formula IIa, V, or VI. More preferably, saidpharmaceutically acceptable derivative is of a preferred compound offormula IIa, V, or VI.

In addition to the compounds of this invention, pharmaceuticallyacceptable derivatives of the compounds of this invention may also beemployed in compositions to treat or prevent the above-identifieddiseases or disorders.

A “pharmaceutically acceptable derivative” means any pharmaceuticallyacceptable salt, ester, salt of an ester or other derivative of acompound of this invention which, upon administration to a recipient, iscapable of providing, either directly or indirectly, a compound of thisinvention or an inhibitorily active metabolite or residue thereof. Themethods for preparing salts or esters of a compound of this inventionare known to one of skill in the art. Particularly favored derivativesare those that increase the bioavailability of the compounds of thisinvention when such compounds are administered to a patient (e.g., byallowing an orally administered compound to be more readily absorbedinto the blood) or which enhance delivery of the parent compound to abiological compartment (e.g., the brain or lymphatic system) relative tothe parent species.

Pharmaceutically acceptable derivatives of the compounds of thisinvention include, without limitation, esters, amino acid esters,phosphate esters, metal salts and sulfonate esters.

Pharmaceutically acceptable salts of the compounds of this inventioninclude those derived from pharmaceutically acceptable inorganic andorganic acids and bases. Examples of suitable acid salts includeacetate, adipate, alginate, aspartate, benzoate, benzenesulfonate,bisulfate, butyrate, citrate, camphorate, camphorsulfonate,cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate,formate, fumarate, glucoheptanoate, glycerophosphate, glycolate,hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide,hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate, malonate,methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oxalate,palmoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate,pivalate, propionate, salicylate, succinate, sulfate, tartrate,thiocyanate, tosylate and undecanoate. Other acids, such as oxalic,while not in themselves pharmaceutically acceptable, may be employed inthe preparation of salts useful as intermediates in obtaining thecompounds of the invention and their pharmaceutically acceptable acidaddition salts.

Salts derived from appropriate bases include alkali metal (e.g., sodiumand potassium), alkaline earth metal (e.g., magnesium), ammonium and N⁺(C₁₋₄ alkyl)₄ salts. This invention also envisions the quaternization ofany basic nitrogen-containing groups of the compounds disclosed herein.Water or oil-soluble or dispersible products may be obtained by suchquaternization.

It should also be understood that a specific dosage and treatmentregimen for any particular patient will depend upon a variety offactors, including the activity of the specific compound employed, theage, body weight, general health, sex, diet, time of administration,rate of excretion, drug combination, and the judgment of the treatingphysician and the severity of the particular disease being treated. Thedosage of compound will also depend upon which particular compound is inthe composition

The compounds of this invention are inhibitors of JNK, Src, or Lckkinase as determined by enzymatic assay. Accordingly, these compoundsare useful for treating JNK-, Src-, or Lck-mediated diseases orconditions.

Another aspect of this invention relates to a method for treating aJNK-, Src-, or Lck-mediated disease in a patient, which method comprisesadministering to a patient in need thereof, a therapeutically effectiveamount of a compound of formula I, or a pharmaceutically acceptablecomposition comprising said compound. According to a preferredembodiment, the invention relates to administering a compound of formulaIIa, IIb, IIIa, IIIb, IVa, IVb, V, or VI, or a pharmaceuticallyacceptable composition comprising said compound. A more preferredembodiment relates to administering a compound of formula IIa, V, or VI,or a pharmaceutically acceptable composition comprising said compound.

Yet another aspect of this invention relates to a method for lesseningthe severity of a JNK-, Src-, or Lck-mediated disease in a patient,which method comprises administering to a patient in need thereof, atherapeutically effective amount of a compound of formula I, or apharmaceutically acceptable composition comprising said compound.According to a preferred embodiment, the invention relates toadministering a compound of formula IIa, IIb, IIIa, IIIb, IVa, IVb, V,or VI, or a pharmaceutically acceptable composition comprising saidcompound. A more preferred embodiment relates to administering acompound of formula IIa, V, or VI, or a pharmaceutically acceptablecomposition comprising said compound.

The activity of the compounds of this invention as kinase inhibitors maybe assayed in vitro, in vivo or in a cell line. In vitro assays includeassays that determine inhibition of either the kinase activity or ATPaseactivity of activated enzyme, for example JNK, Lck, or Src. Alternate invitro assays quantitate the ability of the inhibitor to bind to JNK,Lck, or Src and may be measured either by radiolabelling the inhibitorprior to binding, isolating the inhibitor/JNK, inhibitor/Lck, orinhibitor/Src complex and determining the amount of radiolabel bound, orby running a competition experiment where new compounds are incubatedwith JNK, Lck, or Src bound to known radioligands. One may use any typeor isoform of JNK, Lck, or Src, depending upon which JNK, Lck, or Srctype or isoform is to be inhibited. The details of the conditions usedfor the enzymatic assays are set forth in the Examples hereinbelow.

The term “JNK-mediated disease” or “condition”, as used herein means anydisease or other deleterious condition in which JNK is known to play arole. Such conditions include, without limitation, inflammatorydiseases, autoimmune diseases, destructive bone disorders, proliferativedisorders, cancer, infectious diseases, neurodegenerative diseases,allergies, reperfusion/ischemia in stroke, heart attacks, angiogenicdisorders, organ hypoxia, vascular hyperplasia, cardiac hypertrophy,thrombin-induced platelet aggregation, and conditions associated withprostaglandin endoperoxidase synthase-2.

Inflammatory diseases which may be treated or prevented by the compoundsof this invention include, but are not limited to, acute pancreatitis,chronic pancreatitis, asthma, allergies, and adult respiratory distresssyndrome.

Autoimmune diseases which may be treated or prevented by the compoundsof this invention include, but are not limited to, glomerulonephritis,rheumatoid arthritis, systemic lupus erythematosus, scleroderma, chronicthyroiditis, Graves' disease, autoimmune gastritis, diabetes, autoimmunehemolytic anemia, autoimmune neutropenia, thrombocytopenia, atopicdermatitis, chronic active hepatitis, myasthenia gravis, multiplesclerosis, inflammatory bowel disease, ulcerative colitis, Crohn'sdisease, psoriasis, or graft vs. host disease.

Destructive bone disorders which may be treated or prevented by thecompounds of this invention include, but are not limited to,osteoporosis, osteoarthritis and multiple myeloma-related bone disorder.

Proliferative diseases which may be treated or prevented by thecompounds of this invention include, but are not limited to, acutemyelogenous leukemia, chronic myelogenous leukemia, metastatic melanoma,Kaposi's sarcoma, multiple myeloma and HTLV-1 mediated tumorigenesis.

Angiogenic disorders which may be treated or prevented by the compoundsof this invention include solid tumors, ocular neovasculization,infantile haemangiomas. Infectious diseases which may be treated orprevented by the compounds of this invention include, but are notlimited to, sepsis, septic shock, and Shigellosis.

Viral diseases which may be treated or prevented by the compounds ofthis invention include, but are not limited to, acute hepatitisinfection (including hepatitis A, hepatitis B and hepatitis C), HIVinfection and CMV retinitis.

Neurodegenerative diseases which may be treated or prevented by thecompounds of this invention include, but are not limited to, Alzheimer'sdisease, Parkinson's disease, amyotrophic lateral sclerosis (ALS),epilepsy, seizures, Huntington's disease, traumatic brain injury,ischemic and hemorrhaging stroke, cerebral ischemias orneurodegenerative disease, including apoptosis-driven neurodegenerativedisease, caused by traumatic injury, acute hypoxia, ischemia orglutamate neurotoxicity.

“JNK-mediated disease” or “condition” also includes ischemia/reperfusionin stroke, heart attacks, myocardial ischemia, organ hypoxia, vascularhyperplasia, cardiac hypertrophy, hepatic ischemia, liver disease,congestive heart failure, pathologic immune responses such as thatcaused by T cell activation and thrombin-induced platelet aggregation.

In addition, JNK inhibitors of the present invention may be capable ofinhibiting the expression of inducible pro-inflammatory proteins.Therefore, other “JNK-mediated diseases” or “conditions” which may betreated by the compounds of this invention include edema, analgesia,fever and pain, such as neuromuscular pain, headache, cancer pain,dental pain and arthritis pain.

The compounds of this invention are also useful as inhibitors ofSrc-family kinases, especially Src. For a general review of thesekinases see Thomas and Brugge, Annu. Rev. Cell Dev. Biol. (1997) 13,513; Lawrence and Niu, Pharmacol. Ther. (1998) 77, 81; Tatosyan andMizenina, Biochemistry (Moscow) (2000) 65, 49. Accordingly, thesecompounds are useful for treating Src-mediated diseases or conditions.

The term “Src-mediated disease” or “condition” as used herein means anydisease or other deleterious condition that is known to be affected bythe activity of one or more Src-family kinases. Such diseases orconditions include hypercalcemia, restenosis, hypercalcemia,osteoporosis, osteoarthritis, symptomatic treatment of bone metastasis,rheumatoid arthritis, inflammatory bowel disease, multiple sclerosis,psoriasis, lupus, graft vs. host disease, T-Cell mediatedhypersensitivity disease, Hashimoto's thyroiditis, Guillain-Barresyndrome, chronic obtructive pulmonary disorder, contact dermatitis,cancer, Paget's disease, asthma, ischemic or reperfusion injury,allergic disease, atopic dermatitis, and allergic rhinitis. Diseasesthat are affected by Src activity, in particular, include hypercalcemia,osteoporosis, osteoarthritis, cancer, symptomatic treatment of bonemetastasis, and Paget's disease.

The term “Lck-mediated disease” or “condition” as used herein means anydisease or other deleterious condition that is known to be affected bythe activity of Lck kinase. Such diseases or conditions includeautoimmune diseases, allergies, rheumatoid arthritis, and leukemia.

A preferred embodiment relates to the method used to treat or prevent aJNK-mediated disease selected from inflammatory diseases, autoimmunediseases, destructive bone disorders, neurodegenerative diseases,allergies, reperfusion/ischemia in stroke, heart attacks, angiogenicdisorders, organ hypoxia, vascular hyperplasia, cardiac hypertrophy, orthrombin-induced platelet aggregation.

Another preferred embodiment relates to the method used to treat orprevent a Src-mediated disease selected from hypercalcemia,osteoperosis, osteoarthritis, or sympomatic treatment of bonemetastasis.

Another preferred embodiment relates to the method used to treat orprevent a Lck-mediated disease selected from autoimmune diseases,rheumatoid arthritis, and leukemia.

Depending upon the particular protein kinase-mediated condition to betreated or prevented, additional therapeutic agents, which are normallyadministered to treat or prevent that condition, may be administeredtogether with the compounds of this invention. For example, in thetreatment of cancer other chemotherapeutic or anti-proliferative agentsmay be combined with the compounds of this invention to treat cancer.These agents include, without limitation, adriamycin, dexamethasone,vincristine, cyclophosphamide, fluorouracil, topotecan, taxol,interferons, and platinum derivatives.

Other examples of agents the compounds of this invention may also becombined with include, without limitation, anti-diabetic agentsincluding insulin or insulin analogues in injectable or inhalation form,glitazones, alpha glucosidase inhibitors, biguanides, insulinsensitizers, and sulfonyl ureaschemotherapeutic agents;anti-inflammatory agents such as corticosteroids, TNF blockers, IL-1 RA,azathioprine, cyclophosphamide, and sulfasalazine; immunomodulatory andimmunosuppressive agents such as cyclosporin, tacrolimus, rapamycin,mycophenolate mofetil, interferons, corticosteroids, cyclophophamide,azathioprine, and sulfasalazine; neurotrophic factors such asacetylcholinesterase inhibitors, MAO inhibitors, interferons,anti-convulsants, ion channel blockers, riluzole, and anti-Parkinsonianagents; agents for treating cardiovascular disease such asbeta-blockers, ACE inhibitors, diuretics, nitrates, calcium channelblockers, and statins; agents for treating liver disease such ascorticosteroids, cholestyramine, interferons, and anti-viral agents;agents for treating blood disorders such as corticosteroids,anti-leukemic agents, and growth factors; and agents for treatingimmunodeficiency disorders such as gamma globulin.

Those additional agents may be administered separately from thecompound-containing composition, as part of a multiple dosage regimen.Alternatively, those agents may be part of a single dosage form, mixedtogether with the compound of this invention in a single composition. Ifadministered as part of a multiple dosage regime, the two active agentsmay be submitted simultaneously, sequentially or within a period of timefrom one another normally within five hours from one another.

The amount of both, the compound and the additional therapeutic agent(in those compositions which comprise an additional therapeutic agent asdescribed above)) that may be combined with the carrier materials toproduce a single dosage form will vary depending upon the host treatedand the particular mode of administration. Preferably, the compositionsof this invention should be formulated so that a dosage of between0.01-100 mg/kg body weight/day of a compound of formula I can beadministered.

In those compositions which comprise an additional therapeutic agent,that additional therapeutic agent and the compound of this invention mayact synergistically. Therefore, the amount of additional therapeuticagent in such compositions will be less than that required in amonotherapy utilizing only that therapeutic agent. In such compositionsa dosage of between 0.01-100 μg/kg body weight/day of the additionaltherapeutic agent can be administered.

The compounds of this invention, or pharmaceutical compositions thereof,may also be incorporated into compositions for coating an implantablemedical device, such as prostheses, artificial valves, vascular grafts,stents and catheters. Vascular stents, for example, have been used toovercome restenosis (re-narrowing of the vessel wall after injury).However, patients using stents or other implantable devices risk clotformation or platelet activation. These unwanted effects may beprevented or mitigated by pre-coating the device with a pharmaceuticallyacceptable composition comprising a kinase inhibitor. Suitable coatingsand the general preparation of coated implantable devices are describedin U.S. Pat Nos. 6,099,562; 5,886,026; and 5,304,121. The coatings aretypically biocompatible polymeric materials such as a hydrogel polymer,polymethyldisiloxane, polycaprolactone, polyethylene glycol, polylacticacid, ethylene vinyl acetate, and mixtures thereof. The coatings mayoptionally be further covered by a suitable topcoat of fluorosilicone,polysaccarides, polyethylene glycol, phospholipids or combinationsthereof to impart controlled release characteristics in the composition.Implantable devices coated with a compound of this invention are anotherembodiment of the present invention.

Each of the aforementioned methods directed to the inhibition of JNK,Lck, or Src, or the treatment of a disease alleviated thereby, ispreferably carried out with a preferred compound of formula I, IIa, V,or VI, as described above. More preferably, each of the aforementionedmethods is carried out with a preferred compound of formula I, IIa, V,or VI, and most preferably with a compound of formula IIa, V, or VI.

In order that the invention described herein may be more fullyunderstood, the following examples are set forth. It should beunderstood that these examples are for illustrative purposes only andare not to be construed as limiting this invention in any manner.

EXAMPLES

As used herein,the term “R_(t)(min)” refers to the HPLC retention time,in minutes, associated with the compound using the HPLC methodspecified. Unless otherwise indicated, the HPLC methods utilized toobtain the reported retention times are as follows:

Method-A:

Column: YMC ODS-AQ, 30×100 mm

Gradient: 10%→90% CH₃CN/water (0.2% TFA) over 5 minutes

Flow rate: 1 mL/minute

Method-B:

Column: YMC ODS-AQ, 30×100 mm

Gradient: 10%→90% CH₃CN/water (0.01% TFA)

Flow rate: 1 mL/minute

Detection: 210, 220, 254, 280, and 300 nm.

Method-C:

Column: Lightning, 2.1×50 mm

Gradient: 100% water (0.1% TFA)→100%

CH₃CN (0.1% TFA)

Flow rate: 0.8 mL/minute

Example 1

5-Acetyl-2-methylsulfanyl-4-(4-chloro-phenyl)-thiophene-3-carbonitrile

3-(4-chloro-phenyl)-3-oxo-propionitrile (5 mmol) was added to asuspension of K₂CO₃ (3 equivalents, 15 mmol) in DMF (4.5 mL) and allowedto stir at room temperature. After 10 minutes, CS₂ (1.25 equivalents,7.5 mmol) was added in one portion and the resulting mixture stirred atroom temperature for an additional 10 minutes then a solution of1-chloro-propan-2-one (1.0 equivalent, 5 mmol) in DMF (5 mL) was added.After 1 hour, a solution of MeI (1.1 equivalents, 5.5 mmol) in DMF (2mL) was added in a dropwise fashion then, after 30 minutes, the mixturewas poured onto water and the resulting mixture was stirred vigorouslyfor 12-16 hours to afford a suspension of the desired product. The crudeproduct was isolated by filtration.

Example 2

2-Methylsulfanyl-5-(2-phenylamino-pyrimidin-4-yl)-4-(4-chloro-phenyl)-thiophene-3-carbonitrile(IIa-10) and2-Ethoxy-5-(2-phenylamino-pyrimidin-4-yl)-4-(4-chloro-phenyl)-thiophene-3-carbonitrile(IIa-11)

The crude5-acetyl-2-methylsulfanyl-4-(4-chloro-phenyl)-thiophene-3-carbonitrile(1 mmol) was combined with Brederick's reagent (150 μL) in THF (10 mL)and allowed to stir at room temperature for 12-16 hours. The reactionmixture was concentrated and crude concentrate was dissolved in EtOH (10mL). N-Phenyl guanidine (1.2 equivalent, 1.2 mmol) and sodium acetate (1equivalent, 1 mmol) were added to the ethanolic solution and theresulting mixture was heated to 90° C. for 4 hours. The reaction mixturewas concentrated then the residue was dissolved in ethyl acetate. Theresulting organic solution was washed sequentially with water and brine.The organic layer was concentrated then the crude residue was purifiedby preparatory HPLC to afford compounds IIa-10 and IIa-11.

Example 3

5-Acetyl-2-methylsulfanyl-4-(4-methyl-phenyl)-thiophene-3-carbonitrile

3-(4-Methyl-phenyl)-3-oxo-propionitrile (5 mmol) was added to asuspension of K₂CO₃ (3 equivalents, 15 mmol) in DMF (4.5 mL) and allowedto stir at room temperature. After 10 minutes, CS₂ (1.25 equivalents,7.5 mmol) was added in one portion and the resulting mixture stirred atroom temperature for an additional 10 minutes. A solution of1-chloro-propan-2-one (1.0 equivalent, 5 mmol) in DMF (5 mL) was added.After 1 hour, a solution of MeI (1.1 equivalents, 5.5 mmol) in DMF (2mL) was added in a dropwise fashion. After 30 minutes, the mixture waspoured onto water and the resulting mixture was stirred vigorously for12-16 hours to afford a suspension of the desired product. The crudeproduct was isolated by filtration.

Example 4

2-Methylsulfanyl-5-(2-phenylamino-pyrimidin-4-yl)-4-p-tolyl-thiophene-3-carbonitrile(IIa-14) and2-Ethoxy-5-(2-phenylamino-pyrimidin-4-yl)-4-p-tolyl-thiophene-3-carbonitrile(IIa-15)

The crude5-acetyl-2-methylsulfanyl-4-(4-methyl-phenyl)-thiophene-3-carbonitrile(1 mmol) was combined with Brederick's reagent (150 μL) in THF (10 mL)and allowed to stir at room temperature for 12-16 hours. The reactionmixture was concentrated and crude concentrate was dissolved in EtOH (10mL). N-Phenyl guanidine (1.2 equivalent, 1.2 mmol) and sodium acetate (1equivalent, 1 mmol) were added to the ethanolic solution and theresulting mixture was heated to 90° C. for 4 hours. The reaction mixturewas concentrated then the residue was dissolved in ethyl acetate. Theresulting organic solution was washed sequentially with water and brine.The organic layer was concentrated then the crude residue was purifiedby preparative HPLC to afford compounds IIa-14 and IIa-15.

Example 5

3-Methyl-5-methylsulfanyl-4-(2-phenylamino-pyrimidin-4-yl)-thiophene-2-carbonitrile(IIa-37)

4-(3-Dimethylamino-acryloyl)-3,4-dimethyl-5-methylsulfanyl-4,5-dihydro-thiophene-2-carbonitrile(0.2 mmol) was combined with N-phenyl-guanidine (0.2 mmol) inacetonitrile (0.25 mL) and the resulting mixture heated at reflux for 24hours. The reaction mixture was diluted with methanol (3mL) and theresulting slurry filtered and the solid was washed with methanol (2 mL)then dried under nitrogen to afford IIa-37. M+H=507.1, HPLC: R_(t)=6.196minutes, ¹H NMR (500 Mz, DMSO) 9.77(s, 1H), 8.25(d, 1H, J=5.3 Hz),7.69(s, 1H), 7.48(m, 2H), 7.48(m, 4H), 7.41(t, 2H, J=7.3 Hz), 7.34(t,1H, J=7.2 Hz), 7.25(d, 1H, J=8.3 Hz), 7.21 (t, 1H, J=8.0 Hz), 6.67(d,1H, J=6.7 Hz), 6.01(d, 1H, J=5.3 Hz), 5.11(s, 2H), 3.32(s, 3H, CH3),2.68(s, 3H, CH3).

Example 6

3-[4-(4-cyano-5-methylsulfanyl-3-phenyl-thiophen-2-yl)-pyrimidin-2-ylamino]-benzenesulfonamide(IIa-38)

M+H=480.1, HPLC R_(t)=5.018 minutes, ¹H NMR (500 Mz, DMSO) 10.12(s, 1H),8,64(s, 1H), 8.28(d, 1H, J=5.3 Hz), 7.69(d, 1H, J=7.7 Hz), 7.60-7.58(m,3H), 7.51(d, 1H, J=7.7 Hz), 7.49-7.47(m, 4H), 7.32(br s, 1H), 7.00(br s,1H), 6.06(d, 1H, J=5.3 Hz), 2.90(s, 3H).

Example 7

2-(4-Hydroxy-benzylsulfanyl)-4-phenyl-5-(2-phenylamino-pyrimidin-4-yl)-thiophene-3-carbonitrile(IIa-39)

M+H=493.2, HPLC: R_(t)=5.812 minutes, ¹NMR (500 Mz, DMSO) 9.75(s, 1H),8.24(d, 1H, J=5.2 Hz), 7.73(d, 2H, J=8.0 Hz), 7.57(m, 3H), 7.44(m, 2H),7.34(t, 2H, J=7.9 Hz), 7.26(d, 2H, J=8.5 Hz), 7.01(t, 1H, 7.2 Hz),6.74(d, 2H, J=8.5 Hz), 6.01(d, 1H, J=5.2 Hz), 4.43(s, 2H).

Example 8

5-[2-(3-Hydroxy-phenylamino)-pyrimidin-4-yl]-2-methylsulfanyl-4-phenyl-thiophene-3-carbonitrile(IIa-40)

M+H=417.1, HPLC: R_(t)=4.682 minutes, ¹NMR (500 Mz, DMSO) 9.62(s, 1H),9.26(s, 1H), 8.22(d, 1H, J=5.3 Hz), 7.58(m, 3H), 7.48(m, 2H), 7.30(t,1H, J=1.9 Hz), 7.16(d, 1H, J=8.1 Hz), 7.07(t, 1H, J=8.0 Hz), 6.40(dd,1H, J=7.9, 2.2 Hz), 5.99(d, 1H, J=5.2 Hz), 2.84(s, 3H).

Example 9

5-[2-(3-Benzyloxy-phenylamino)-pyrimidin-4-yl]-2-hydroxy-4-phenyl-thiophene-3-carbonitrile(IIa-42)

M+H=477.2, HPLC: Rt=4.530 minutes, ¹NMR (500 Mz, DMSO) 10.13(s, 1H),7.55(m, 4H), 7.48-7.38(m, 5H), 7.34(t, 1H, J=7.2 Hz), 7.29(t, 1H, J=8.2Hz), 7.12(d, 1H, J=7.8 Hz), 6.79(dd, 1H, J=8.2, 2.1 Hz), 5.63(d, 1H,J=7.1 Hz), 5.18(s, 2H).

Example 10

5-[2-(3-Hydroxy-phenylamino)-pyrimidin-4-yl]-2-methylsulfanyl-4-(3-pyridin-4-yl-phenyl)-thiophene-3-carbonitrile(IIa-105)

To 2 ml DME in a 10 ml tube was added I (50 mg, 0.10 mmol),4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)pyridine (25 mg, 0.12mmol) and tetrakis(triphenylphosphine)palladium(0) (11.5 mg, 0.01 mmol).The mixture was stirred at ambient temperature under nitrogen for 30minutes then 0.3 ml saturated NaHCO₃ aqueous solution was added to thereaction mixture and the reaction was stirred at 45° C. in a sealed tubefor 6 hours. The reaction was cooled to ambient temperature then wasdiluted with 2 ml water. The mixture was extracted with ethyl acetate (5ml×3) and combined organic layers were dried over Na₂SO₄. The solventwas removed under reduced pressure and the residue purified bychromatography using ethyl acetate and hexane (1:1) on silica gel toafford 23 mg of the desired compounds in 47% yield. ¹NMR δ (Acetone-d6):8.96 (s, 1H) , 8.92(d, 2H), 8.20(m, 5H), 7.88(dd, 1H), 7.80(dd, 1H),7.53(d, 1H), 7.32(d, 1H), 7.22(dd, 1H), 6.56(d, 1H), 6.26(d, 1H),2.92(s, 3H)

Example 11

We have prepared other compounds of formula IIa by methods substantiallysimilar to those described in the above Examples 1-10 and thoseillustrated in Scheme I. The characterization data for these compoundsis summarized in Table 9 below and includes LC/MS (observed) and ¹HNMRdata. “Y” designates ¹HNMR data was obtained and found to be consistantwith the assigned structure. Compound numbers correspond to the compoundnumbers listed in Table 1.

TABLE 9 Characterization Data for Selected Compounds of Formula IIaCompound No M + 1 (obs) R_(t) Method ¹H NMR IIa-1  368.04 — — Y IIa-2 358.01 — — Y IIa-3  392.03 — — Y IIa-4  338.06 — — — IIa-5  358.01 — — YIIa-6  354.06 — — — IIa-7  451.00 — — — IIa-8  417.00 — — — IIa-9 413.01 — — — IIa-10 — 4.935 B Y IIa-11 — 4.732 B Y IIa-12 468.00 4.969 BY IIa-13 466.00 4.784 B Y IIa-14 414.01 4.888 B Y IIa-15 412.01 4.627 BY IIa-16 324.05 — — Y IIa-37 507.1 6.193 B Y IIa-38 479.9 5.018 B YIIa-39 493.0 5.812 B Y IIa-40 447.1 4.682 B Y IIa-42 477.2 4.530 B YIIa-43 481.3 5.10 A Y IIa-44 586.2 4.18 A Y IIa-45 513.0 5.82 A Y IIa-46407.0 6.28 A Y IIa-47 479.0 5.51 A Y IIa-48 403.1 4.42 A Y IIa-49 4762.62 A Y IIa-50 446.08 2.99 A Y

Example 12

4-Acetyl-3-methyl-5-methylsulfanyl-thiophene-2-carbonitrile

To a slurry of 2,4-pentanedione (20 mmol) and K₂CO₃ (3 equivalents, 60mmol) in DMF (10 mL) was added CS₂ (1.2 equivalents, 24 mmol) and theresulting mixture stirred for 10 minutes. The mixture was cooled to 0°C. then chloroacetonitrile (1.0 equivalent, 20 mmol) was added and thereaction stirred 1 hour then warmed to room temperature and stirred foran additional 2 hours. The reaction mixture was again cooled to 0° C.and methyl iodide (1.05 equivalents, 22 mmol) was added slowly. Theresulting mixture was warmed to room temperature. After 12 hours, waterwas added and the resulting suspension was stirred overnight. Thedesired product was isolated by filtration after washing with water.

Example 13

4-(3-Dimethylamino-acryloyl)-3-methyl-5-methylsulfanyl-thiophene-2-carbonitrile

To a solution of4-acetyl-3-methyl-5-methylsulfanyl-thiophene-2-carbonitrile (10 mmol) inacetonitrile (5 mL) was added DMF-DMA (2 mL) and the resulting mixtureheated at reflux for 18 hours. The reaction was concentrated and theresidue triturated with diethyl ether (20 mL). The suspension wasfiltered and washed with ether to afford the desired product as a yellowsolid.

Example 14

3-Methyl-5-methylsulfanyl-4-(2-phenylamino-pyrimidin-4-yl)-thiophene-2-carbonitrile(IIb-7)

To a solution of4-(3-dimethylamino-acryloyl)-3-methyl-5-methylsulfanyl-thiophene-2-carbonitrile(0.2 mmol) in acetonitrile (0.25 mL) was added N-phenyl guanidine (1equivalent, 0.2 mmol) and the reaction heated at reflux for 24 hours.The resulting mixture was cooled to room temperature then diluted withmethanol (3 mL). The resulting slurry was filterred washed with methanolto afford IIb-7.

Example 15

3-Dimethylamino-1-[4-methyl-2-methylsulfanyl-5-(4-methyl-thiazol-2-yl)-thiophen-3-yl]-propenone

To a solution of1-[4-methyl-2-methylsulfanyl-5-(4-methyl-thiazole-2-yl)-thiophen-3-yl]-ethanone(10 mmol, Maybridge Chemicals) in acetonitrile (5 mL) was added DMF-DMA(2 mL) and the resulting mixture heated at reflux for 18 hours. Thereaction was concentrated and the residue triturated with diethyl ether(20 mL). The suspension was filtered and washed with ether to afford thedesired product as a yellow solid.

Example 16

{4-[4-methyl-2-methylsulfanyl-5-(4-methyl-thiazol-2-yl)-thiophen-3-yl]-pyrimidin-2-yl}-phenylamine(IIb-1)

To a solution of3-dimethylamino-1-[4-methyl-2-methylsulfanyl-5-(4-methyl-thiazol-2-yl)-thiophen-3-yl]-propenone(0.2 mmol) in acetonitrile (0.25 mL) was added N-phenyl guanidine (1equivalent, 0.2 mmol) and the reaction heated at reflux for 24 hours.The resulting mixture was cooled to room temperature then diluted withmethanol (3 mL). The resulting slurry was filterred washed with methanolto afford IIb-1.

Example 17

We have prepared other compounds of formula IIb by methods substantiallysimilar to those described in the above Examples 12-16 and thoseillustrated in Schemes II and III. The characterization data for thesecompounds is summarized in Table 10 below and includes ah LC/MS(observed) data. Compound numbers correspond to the compound numberslisted in Table 2.

TABLE 10 Characterization Data for Selected Compounds of Formula IIbCompound No M + 1 (obs) Compound No M + 1 (obs) IIb-1 410.9 IIb-7  339.1IIb-2 428.9 IIb-8  357.2 IIb-3 445.9 IIb-9  374.1 IIb-4 444.9 IIb-10373.1 IIb-5 440.9 IIb-11 369.2 IIb-6 516.9 IIb-12 445.1

Example 18

Resin-bound4-(3-chloro-phenyl)-2-(4-resin-oxy-benzylsulfanyl)-5-propionyl-thiophene-3-carbonitrile

To a stirring slurry of 3-chlorobenzoylacetonitrile (Maybridge, 1equivalent, 15 mmol) and LiOH-H₂O (3 equivalents, 45 mmol) in DMF (30mL) was added CS₂ (1.2 equivalents, 18 mmol) and the resulting mixturestirred for 10 minutes at 0° C. 1-bromo-2-butanone (1.0 equivalent, 15mmol) was added to the mixture at 0˜5° C. and stirred for one hour.Bromo-Wang resin (Novabiochem, 3 g, 3.9 mmol, 1.3 mmol/g loading) wasthen added at 0˜5° C. and the resulting mixture was stirred at roomtemperature for 12 hours. The solvent was removed by filtration and theresin rinsed with DMF (30 mL,×3), THF (30 mL,×3), water (30 mL×3), DMF(30 mL×3), DCM (30 mL×3) and diethyl ether (30 mL×3) then dried undernitrogen to afford the title compound.

Example 19

Resin bound4-(3-chloro-phenyl)-5-(3-dimethylamino-2-methyl-acryloyl)-2-(4-hydroxy-benzylsulfanyl)-thiophene-3-carbonitrile

The above resin bound compound formed in Example 18 above was slurriedin dry THF (20 mL) and treated with DMF-DMA (2 mL) then the resultingmixture was heated at 60° C. for 18 hours. The solvent was removed byfiltration and the resin was rinsed with THF (30 mL×5), DCM (30 mL×3),and diethyl ether (30 mL×3) then dried under nitrogen to afford thetitle compound.

Example 20

Resin bound 4-(3-chloro-phenyl)-2-(4-resin boundedhydroxy-benzylsulfanyl)-5-(5-methyl-2-phenylamino-pyrimidin-4-yl)-thiophene-3-carbonitrile

The resin bound compound formed in Example 19 above (1 g) andN-phenyl-guanidine (4 mmol) were combined in dry THF (10 mL). Theresulting mixture was heated to reflux for 24 hours then the solvent wasremoved by filtration and the resin rinsed with THF (30 mL×5), DMF(30mL×3), DCM (30 mL×3), and diethyl ether (30 mL×3) then dried undernitrogen. The resin was treated once again with N-phenyl-guanidine (4mmol) in dry THF (10 mL) for 24 hours. Solvent was again removed byfiltration and the resin rinsed with THF (30 mL×5), DMF (30 mL×3), DCM(30 mL×3), and diethyl ether (30 mL×3) then dried under nitrogen toafford the title compound.

Example 21

4-(3-chloro-phenyl)-2-(4-hydroxy-benzylsulfanyl)-5-(5-methyl-2-phenylamino-pyrimidin-4-yl)-thiophene-3-carbonitrile(V-1)

The resin bound compound formed at Example 20 above(100 mg, 0.1 mmol)was slurried in DCM (1 mL) and treated with TFA (1 mL) and one drop ofwater for 3 hours. The resin was filtered and washed with DCM (2 mL×3).The resin was then treated with 50% TFA in DCM-water (1 mL:1 mL: 0.02mL). The mixture was set aside for 14 hours then filtered. The resin waswashed with dichloromethane (2 mL×3), the combined organic layers wereconcentrated, and the crude product was purified by preparative HPLC[YMC ODS-AQ column, gradient 10%→90% B (solvent A: 0.01% TFA in water;solvent B: 0.01% TFA in CH₃CN) over 6.5 minutes at 1 mL/min] to affordcompound V-1 (10 mg). [M+H]=541.1.

Example 22

We have prepared other compounds of formula V by methods substantiallysimilar to those described in the above Examples 18-21 and thoseillustrated in Scheme IV. The characterization data for these compoundsis summarized in Table 11 below and includes HPLC, LC/MS (observed) and¹NMR data.

¹H NMR data is summarized in Table 11 below wherein “Y” designates ¹HNMR data is available and was found to be consistant with structure.

Compound numbers correspond to the compound numbers listed in Table 7.

TABLE 11 Characterization Data for Selected Compounds of Formula VCompound No M + 1 (obs) R_(t)(min) Method ¹H NMR V-4  521.2 6.063 B YV-5  494.1 5.114 B Y V-6  431.1 4.97 B Y V-7  460.1 5.930 B Y V-8  430.13.987 B — V-9  489.1 4.711 C Y V-10 499.0 4.024 C Y V-11 541.1 — — —V-13 445.1 5.197 B — V-14 430.1 4.115 B — V-15 522.2 6.75 B Y V-16 416.13.67 A Y V-17 446.1 3.69 A Y V-18 476.1 3.73 A Y V-19 494.0 4.18 A YV-20 547.2 2.51 A Y V-21 474.1 3.74 A Y V-22 451.0 3.44 A Y V-23 430.13.61 A Y V-24 432.1 5.47 B Y V-25 406.1 4.893 B — V-26 421.1 4.749 B YV-27 511.0 6.608 B Y V-28 450.0 5.923 B —

Example 23

3-oxo-3-pyridin-3-yl-propionitrile

To a solution of ethyl nicotinate (30.24 g, 0.20 mol) in toluene(anhydrous, 200 mL) was added NaH (18.64 g, 0.47 mol, 60% in mineraloil). The resulting suspension was heated at 90° C. and acetonitrile(anhydrous, 24.74 ml, 0.47 mol) was added into this suspension viasyringe under nitrogen and the reaction was heated at 90° C. overnight.The reaction mixture was cooled to ambient temperature and the resultingsolid material was collected by filtration. The crude product was driedin vacuo and used directly in the next step. For analytical purposes,the solid was dissolved in water. The pH was adjusted to 5 by aqueousHCl and the solution extracted with DCM. The organic layer was driedover Na₂SO₄ and the solvent was removed in vacuo to afford a sample ofthe product for characterization. ¹H NMR(CDCl₃): δ9.2(s, 1H), 8.8(d,1H), 8.2(d, 1H), 7.5(dd, 1H), 4.1 (s, 2H)

Example 24

1-(4-Methyl-5-methylsulfanyl-3-pyridin-3-yl-thiophen-2-yl)-propan-1-one

To a solution of 3-oxo-3-pyridin-3-yl-propionitrile (11.9 mmol) in DMF(100 mL) was added CS₂ (0.90 ml, 15.0 mmol) at 0° C. The reaction wasstirred at 0° C. for 45 minutes under nitrogen then 1-bromo-butan-2-one(1.79 g, 11.8 mmol) was added via a syringe at 0° C. The reaction wasstirred for another 30 minutes then methyl iodide (1.69 g, 11.8 mmol)was added via syringe at 0° C. The resulting mixture was stirred foranother 30 minutes then warmed to ambient temperature and poured intosaturated aqueous NH₄Cl solution (300 mL). The solution was extractedwith ethyl acetate (200 mL×3) and the combined organic layers were driedover Na₂SO₄ then concentrated in vacuo. Purification by chromatography(Silica Gel, 3:1 hexanes:ethyl acetate) afforded 1.05 g desired product.¹H NMR (CDCl3): δ8.8(d, 1H), 8.6 (s, 1H), 7.8 (d, 1H), 7.5 (dd, 1H),2.8(s, 3H), 2.3 (q, 2H), 1.0 (t, 3H).

Example 25

5-(3-Dimethylamino-2-methyl-acryloyl)-2-methylsulfanyl-4-pyridin-3-yl-thiophene-3-carbonitrile

A solution of1-(4-methyl-5-methylsulfanyl-3-pyridin-3-yl-thiophen-2-yl)-propan-1-one(1.0 g, 3.48 mmol) in DMF-DMA (10 mL) in a 50 ml sealed tube was stirredat 90° C. overnight. The reaction was cooled to ambient temperature andthe excess dimethylformamide dimethyl acetal was removed in vacuo. Thepurification of the crude by chromatography, eluting with ethyl acetate,afforded 964 mg (81%) desired product. ¹H NMR (CDCl₃): δ8.6(m, 2H) , 7.7(d, 1H) , 7.3 (d, 1H), 6.7 (s, 1H),2.8 (s, 6H), 2.7 (s, 3H), 1.9 (s,3H).

Example 26

N-(3-Benzyloxy-phenyl)guanidine

To a suspension of 3-benzyloxyphenylamine (20.0 g, 100.35 mmol) in1,4-dioxane (150 mL) was added cyanamide (7.39 g, 175.95 mmol followedby 4M HCl in 1,4-dioxane (44 ml, 176.00 mmol).

The resulting suspension was heated at 80° C. overnight then cooled toambient temperature and 6N NaOH (35 ml, 210.00 mmol) was added. Thevolume of solution was reduced to 50 ml in vacuo and the resultingprecipitate was collected by filtration. The solid product was driedunder vacuum overnight to afford23.8 g in 98.4% yield. ¹H NMR (MeOH-d4)δ6.4-7.5 (m, 9H), 5.1 (s, 2H).

Example 27

N-(3-Hydroxy-phenyl)-guanidine (5)

To a solution of N-(3-benzyloxy-phenyl)guanidine (5.0 g, 20.6 mmol) inEtOH (150 mL) was added palladium on carbon (10%, wet, 50% water, 0.5g)). The mixture was stirred under hydrogen atmosphere overnight. Thereaction was filtered through a plug of celite and the filtrateconcentrated in vacuo. with toluene azeotroping to affored the desiredmaterial (2.83 g, 91%). ¹H NMR (MeOH-d4) δ6.4-7 (m, 4H)

Example 28

5-[5-(3-Hydroxy-phenylamino)-2-methyl-phenyl]-2-methylsulfanyl-4-pyridin-3-yl-thiophene-3-carbonitrile(V-24)

To a solution of5-(3-dimethylamino-2-methyl-acryloyl)-2-methylsulfanyl-4-pyridin-3-yl-thiophene-3-carbonitrile(45 mg, 0.13 mmol) in CH₃CN (anhydrous, 2 mL) was addedN-(3-hydroxy-phenyl)-guanidine (38 mg, 0.16 mmol). The reaction wasstirred at 90° C. in a sealed tube overnight then cooled to ambienttemperature and the solvent removed in vacuo. The crude product waspurified by chromatography (Silica Gel, 2:1 hexanes:ethyl acetate) toafford compound V-24 (51 mg, 75%). ¹H NMR (CDCl₃): δ8.6(d, 1H), 8.5 (s,1H), 8.2 (s, 1H), 7.8 (d, 1H),7.6 (s, 1H), 7.5-7.1 (m, 8H), 7.0 (d, 1H),6.7 (d, 1H), 5.1 (s, 2H), 2.6 (s, 3H), 1.5 (s, 3H).

Example 29

5-{5-[3-(3-Hydroxy-propoxy)-phenylamino]-2-methyl-phenyl}-2-methylsulfanyl-4-pyridin-3-yl-thiophene-3-carbonitrile(VI-29)

To a solution of5-[5-(3-hydroxy-phenylamino)-2-methyl-phenyl]-2-methylsulfanyl-4-pyridin-3-yl-thiophene-3-carbonitrile(330 mg, 0.77 mmol) in DMF (anhydrous, 5 mL) was added3-bromo-propan-1-ol (214 mL, 1.54 mmol). The reaction was stirred at 70°C. in the presence of excess K₂CO₃ overnight then cooled to ambienttemperature and partitioned between water and DCM. The mixture wasextracted with DCM (30 ml×3) and the combined organic layers were driedover Na₂SO₄ and concentrated in vacuo. Purification by chromatography(2% MeOH-DCM) afforded desired product (370 mg, 99%). ¹H NMR (CDCl3):δ8.6(d, 1H), 8.5 (d, 1H), 8.1 (s, 1H), 7.7 (d, 1H), 7.4 (s, 1H), 7.3-7.2(m, 3H), 7.0 (d, 1H), 6.6 (d, 1H), 4.1 (t, 2H), 3.9 (t, 2H), 2.7 (s,3H), 2.0 (m, 2H), 1.5 (s, 3H)

Example 30

Methanesulfonic acid3-{3-[3-(4-cyano-5-methylsulfanyl-3-pyridin-3-yl-thiophene-2-yl)-4-methyl-phenylamino]-phenoxyl}-propylester

To a solution of5-{5-[3-(3-hydroxy-propoxy)-phenylamino]-2-methyl-phenyl}-2-methylsulfanyl-4-pyridin-3-yl-thiophene-3-carbonitrile(370 mg, 0.76 mmol) in DCM (10 mL) was added Et₃N (155 mg, 0.15 mmol)followed by MsCl (130 mg, 1.15 mmol). The resulting mixture was stirredat ambient temperature for 10 min under nitrogen then poured into 20 mlwater and extracted with DCM (20 ml×3). The combined organic layers weredried over Na₂SO₄ and concentrated in vacuo. The desired product wasobtained (391 mg, 91%) and confirmed by LC/MS (Calculated: 565.12;observed: 565.1+1, ES+).

Example 31

5-{5-[3-(3-Dimethylamino-propoxy)-phenylamino]-2-methyl-phenyl}-2-methylsulfanyl-4-pyridin-3-yl-thiophene-3-carbonitrile(VI-18)

To a solution of methanesulfonic acid3-{3-[3-(4-cyano-5-methylsulfanyl-3-pyridin-3-yl-thiophene-2-yl)-4-methyl-phenylamino]-phenoxyl}-propylester (20 mg, 0.035 mmol) in DCM (1 mL) was added excess dimethylamineand excess triethylamine. The reaction was stirred at ambienttemperature for 3 hours then water (2 ml) was added and the productextracted with DCM (3 mL×3). The combined organic layer wereconcentrated to afford the crude product as an oil. Purification bychromatography (2% MeOH—CH₂Cl₂) afforded the desired product (17 mg,95%). ¹H NMR (CDCl3): δ8.6(d, 1H), 8.5 (s, 1H), 8.1 (s, 1H), 7.7 (d,1H), 7.4 -7.0 (m, 4H), 7.0(d, 1H), 6.5 (d, 1H), 4.1 (t, 2H), 2.9 (m,2H), 2.7 (s, 3H), 2.6 (s, 6H), 2.1 (m, 2H), 1.6 (s, 3H).

Example 32

2-cyano-3-cyclohexyl-3-oxo-propionic acid ethyl ester

To a solution of ethyl cyanoacetate in acetonitrile was added MgCl₂ andEt₃N at 0° C. The resulting suspension was stirred at 0° C. for 30minutes then cyclehexanecarbonyl chloride was added over 20 minutes andthe reaction mixture stirred at 0° C. for 1 hour. The reaction mixturewas partitioned between 1N HCl and ether, the organic phase was driedwith MgSO₄ then concentrated under reduced pressure.

Example 33

3-cyclohexyl-3-oxo-propionitrile

A solution of 2-cyano-3-cyclohexyl-3-oxo-propionic acid ethyl ester inDMSO:water (10:1) was stirred at 120° C. for 2 hours. The reactionmixture was partitioned between ether and 1N HCl, the organic phase wasdried with MgSO₄ and the solvent removed under reduced pressure.

Example 34

5-Acetyl-4-cyclohexyl-2-methylsulfanyl-thiophene-3-carbonitrile

To a solution of 3-cyclohexyl-3-oxo-propionitrile in DMF:CS₂ (1:1) wasadded K₂CO₃ at 0° C. and the reaction mixture stirred at ambienttemperature for 2 hours. To the resulting reaction mixture was addedchloroacetone (1 equivalent) and the reaction mixture stirred at ambienttemperature for 3 hours. Iodomethane was then added and the resultingmixture was stirred at ambient temperature overnight. The reactionmixture was partitioned between ethyl acetate and brine, the organicphase dried with MgSO₄ and the solvent was removed under reducedpressure. The crude product was purified by silica gel chromatography.

Example 35

5-[2-(3-Nitrophenylamino)-5-methyl-pyrimidin-4-yl]-4-cyclohexyl-2-methylsulfanyl-thiophene-3-carbonitrile(V-7)

To a solution of4-cyclohexyl-5-(3-dimethylamino-2-methyl-acryloyl)-2-methylsulfanyl-thiophene-3-carbonitrile(0.5 g, 1.7 mmol) in toluene (3 ml) was added DMF-DMA and the reactionwas heated at 90° C. overnight resulting in conversion to product asdetermined by TLC (40% EtOAc:hexanes). The crude product was combinedwith DMF (10 ml) and 3-Nitrophenylguanidine (0.5 g, 2.78 mmol) in asealed tube and heated at 120° C. overnight resulting in conversion toproduct as determined by TLC (40% EtOAc:hexanes). The reaction waspartitioned between EtOAc and water, the organics were dried over sodiumsulfate then concentrated in vacuo. The crude product was purified bychromatography (silica gel, 10% EtOAC:hexanes) to afford 0.334 g (3.05umol) of V-7 in 42% yield for 2 steps. ¹H NMR (CDCl₃) δ1.0-2.0 (m, 10H),2.1 (s, 3H), 2.6(s, 3H), 7.4(m, 1H), 7.7 (d, 1H), 7.8 (d, 1H), 8.3 (s,1H), 8.7 (s, 1H).

Example 36

5-[2-(3-Aminophenylamino)-5-methyl-pyrimidin-4-yl]-4-cyclohexyl-2-methylsulfanyl-thiophene-3-carbonitrile(V-8)

A suspension of5-[2-(3-Nitrophenylamino)-5-methyl-pyrimidin-4-yl]-4-cyclohexyl-2-methylsulfanyl-thiophene-3-carbonitrile(0.187 g, 402 umol) and 10% palladium on carbon in methanol was stirredunder a hydrogen atmosphere overnight resulting in complete conversionto product as determined by TLC (40% EtOAc:hexanes). The reaction wasfiltered through celite and concentrated in vacuo to afford the desiredamine V-8 (0.186 g, 426 umol) in quantitative yield. ¹NMR (CDCl₃)δ1.0-1.1(m, 10H), 2.1 (s, 3H), 2.6 (s, 3H), 6.2 (d, 1H), 6.8 (d, 1H),6.9-7.0 (m, 3H) , 8.2 (s, 1H).

Example 37

4-cyclohexyl-5-{2-(3-(2-diethylamino-ethylamino)-phenylamino)-5-methyl-pyrimidin-4-yl}2-methylsulfanyl-thiophene-3-carbonitrile(VI-26)

To a solution of5-[2-(3-aminophenylamino)-5-methyl-pyrimidin-4-yl]-4-cyclohexyl-2-methylsulfanyl-thiophene-3-carbonitrile(20 mg, 45.91 micromol) and N,N-diethyl-2-amino-bromoethane.HBr (12 mg,45.91 micromol) in THF was added sodium t-butoxide (9 mg, 91.82micromol) and the eaction was stirred at ambient temperature overnight.TLC indicated conversion to product (10% MeOH:CH₂Cl₂). The reaction waspartitioned between EtOAc and water, the organics were dried over sodiumsulfate then concentrated in vacuo. The crude product was purified bychromatography (silica gel, 5% MeOH:CH₂Cl₂) to afford VI-26 in 33%yield. δ0.95 (bs, 6H), 1.0-1.8 (m, 10H), 2.0 (s, 3H), 2.6 (s, 3H), 2.75(bs, 2H), 3.4 (s, 1H), 3.6 (bs, 2H), 4.0 (bs, 2H), 5.2 (s, 1H), 6.5 (d,1H), 6.55 (s, 1H) , 6.6 (d, 1H) , 7.0 (t, 1H) , 8.1 (s, 1H).

Example 38

3-cyano-4-(3-pyridyl)-5-[2-(3-N-cyclopropylacetamido)-aminophenylamino]-pyrimidin-4-yl-2-thiomethylthiophene (VI-138)

Placed together in a small vial were:5-[2-(3-aminophenylamino)-pyrimidin-4-yl]-3-cyano-4-(3-pyridyl)-2-methylthio-thiophene(25 mg; 60 umol), cyclopropylacetic acid ( 25 mg; 240 umol),1-(3-dimethylaminopropylo-3-ethylcarbodiimide hydrochloride [EDCI] (25mg; 130 umol); and a catalytic amount (10% w/w) ofN-hydroxy-benzatriazole [HOBT]. One milliliter of a 2.5 mM solution ofdiisopropylethylamine in dry DMF was added to the vial and the resultingsolution was stirred for 18 hours at ambient temperature. Reaction wasmonitored by reverse phase HPLC and determined to be complete. Removalof the solvent via high vacuum and subsequent purification viapreparative reverse phase HPLC on C18 silica with a gradient ofwater/acetonitrile containing 0.1% trifluoroacetic acid afforded eightmilligrams of the title compound as a medium yellow powder; a 25.8%yield. ¹NMR in methanol-d4: δ0.25 (mult; 2H), 0.6 (mult; 2H), 1.15(mult; 1H), 2.3 (d; 2H), 2.85 (s; 3H), 6.4 (d; 1H), 7.15 (mult; 1H),7.21 (mult; 2H), 7.85 (mult; 1H), 8.25 (d; 1H), 8.35 (d; 1H), 8.75(mult; 1H), 8.82 (mult; 1H). m/e (ES+) 499.0

Example 39

3-cyano-4-(3-pyridyl)-5-(-2-(3-tetrahydrofuran-3-methyleneoxycarbamoyl)aminophenylamino)-pyrimidin-4-yl-2-thiomethyl thiophene (VI-143)

5-[2-(3-Aminophenylamino)-pyrimidin-4-yl]-3-cyano-4-(3-pyridyl)-2-thiomethylthiophene (50 mg, 120 uM) was suspended/dissolved in 2.0 mL of a 0.25 Msolution of diisopropylethylamine in p-dioxane. Added to thissuspension/solution was (200 uL, 120 uM) of a 0.55 M solution of2-hydroxymethyltetetrahydrofuran chloroformate in dry p-dioxane (Thechloroformate was made previously according to a literature procedureand utilized as a stock solution.). The resulting solution was stirredat ˜35 C. for 4 hrs. The reaction was determined to be complete byanalytical reverse phase HPLC. The solvent was removed under reducedpressure and the resulting residue was dissolved in DMSO. This crudematerial was purified via preparative reverse phase HPLC on C18 silicautilizing a gradient of water/acetonitrile containing 0.1% TFA. Thisresulted in 10 mg of the title compound as a yellow amorphous powder, in15.2% yield. ¹NMR in methanol d4: δ1.75 (mult, 1H), 2.15 (mult; 1H),2.85 (s; 3H), 3.65 (mult; 1H), 3.76 (mult; 1H), 3.86 (mult; 2H), 4.07(mult; 1H), 4.16 (mult; 1H), ), 6.4 (d; 1H), 7.15 (mult; 1H), 7.21(mult; 2H), 7.85 (mult; 1H), 8.25 (d; 1H), 8.35 (d; 1H), 8.75 (mult;1H), 8.82 (mult; 1H) m/e (ES+) 545.

Example 40

We have prepared other compounds of formula VI by methods substantiallysimilar to those described in the above Examples 22-39 and thoseillustrated in Schemes I-VII. The characterization data for thesecompounds is summarized in Table 12 below and includes ¹NMR and HPLCdata.

¹H NMR data is summarized in Table 12 below wherein “Y” designates ¹NMRdata is available and was found to be consistant with structure.Compound numbers correspond to the compound numbers listed in Table 8.

TABLE 12 Characterization Data for Selected Compounds of Formula VICompound No M + 1 R_(t) Method ¹H NMR VI-1  538.16 3.18 A Y VI-2  538.173.28 A Y VI-3  524.14 3.15 A Y VI-4  564.2 3.23 A Y VI-5  578.21 3.23 AY VI-6  564.20 3.24 A Y VI-7  593.20 2.90 A Y VI-8  564.18 3.22 A YVI-9  577.21 2.98 A Y VI-10 549.17 2.76 A Y VI-11 640.19 2.703 A Y VI-12534.18 3.35 A Y VI-13 531.15 3.28 A Y VI-14 545.13 3.36 A Y VI-15 551.153.25 A Y VI-16 566.13 3.35 A Y VI-17 552.11 3.43 A Y VI-18 517.2 4.81 BY VI-19 547.2 4.57 B Y VI-20 547.2 2.51 A Y VI-21 533.2 2.36 A Y VI-22573.2 2.40 A Y VI-23 587.2 4.64 B Y VI-24 573.2 2.47 A Y VI-25 602.24.78 B Y VI-26 535.21 2.62 A Y VI-27 575.20 3.23 A Y VI-28 549.16 3.13 AY

Example 41 2-Ethylamino-4-phenyl-5-propionyl-thiophene-3-carboxylic acidethyl ester

To a stirring slurry of ethyl benzoylacetate (30 mmol) and K₂CO₃ (1.2equivalents, 36 mmol) in DMF (30 mL) was added ethyl thioisocyanate (1.0equivalent, 30 mmol) and stirred for 16 hours. The chloroacetone (1.0equivalent, 30 mmol) was added to the mixture at room temperature andstirred for 3 hours. To the mixture was added water (150 mL) at roomtemperature with vigorous stirring. The solid was collected and washedwith water (30 mL) and hexane (50 mL). The solid was dried undernitrogen pressure.

Example 42

2-Ethylamino-5-[2-(3-methoxy-phenylamino)-pyrimidin-4-yl]-4-phenyl-thiophene-3-carboxylicacid ethyl ester (IIa-90)

This compound was prepared from2-ethylamino-4-phenyl-5-propionyl-thiophene-3-carboxylic acid ethylester using the essentially the same general methods as described above.MS cal:474.17, obs [M+H]=475.1; ¹H NMR(DMSO) 9.44(s, 1H), 8.37(t, J=5.8Hz, 1H), 7.93(d, J=5.5 Hz, 1H), 7.63(s, 1H), 7.45(m, 3H), 7.24(m, 3H),7.17(t, J=8.1 Hz, 1H), 6.53(dd, J=2.4, 8.1 Hz, 1H), 5.54(d(J=5.6 Hz,1H), 3.80(s, 3H), 3.80(q, J=7.2 Hz, 2H), 3.40(quintet, J=7.1 Hz, 2H),1.32(t, J=7.2 Hz, 3H), 0.67(t, J=7.1 Hz, 3H).

The following examples demonstrate how the compounds of this inventionmay be tested as inhibitors of c-Jun-N-terminal, Src, and Lck kinases.

Example 43 Cloning, Expression and Purification of JNK3 Protein

A BLAST search of the EST database using the published JNK3α1 cDNA as aquery identified an EST clone (#632588) that contained the entire codingsequence for human JNK3α1. Polymerase chain reactions (PCR) using pfupolymerase (Strategene) were used to introduce restriction sites intothe cDNA for cloning into the pET-15B expression vector at the NcoI andBamHI sites. The protein was expressed in E. coli. Due to the poorsolubility of the expressed full-length protein (Met 1-Gln 422), anN-terminally truncated protein starting at Ser residue at position 40(Ser 40) was produced. This truncation corresponds to Ser 2 of JNK1 andJNK2 proteins, and is preceded by a methionine (initiation) and aglycine residue. The glycine residue was added in order to introduce anNcoI site for cloning into the expression vector. In addition,systematic C-terminal truncations were performed by PCR to identify aconstruct that give rise to diffraction-quality crystals. One suchconstruct encodes amino acid residues Ser40-Glu402 of JNK3α1 and ispreceded by Met and Gly residues.

The construct was prepared by PCR using deoxyoligonucleotides: 5′GCTCTAGAGCTCCATGGGCAGCAAAAGCAAAGTTGACAA 3′ (forward primer withinitiation codon underlined)(SEQ ID NO:1) and 5′TAGCGGATCCTCATTCTGAATTCATTACTTCCTTGTA 3′ (reverse primer with stop codonunderlined)(SEQ ID NO:2) as primers and was confirmed by DNA sequencing.Control experiments indicated that the truncated JNK3 protein had anequivalent kinase activity towards myelin basic protein when activatedwith an upstream kinase MKK7 in vitro.

E.coli strain BL21 (DE3) (Novagen) was transformed with the JNK3expression construct and grown at 30° C. in LB supplemented with 100μg/ml carbenicillin in shaker flasks until the cells were in log phase(OD₆₀₀˜0.8). Isopropylthio-β-D-galactosidase (IPTG) was added to a finalconcentration of 0.8 mM and the cells were harvested 2 hours later bycentrifugation.

E. coli cell paste containing JNK3 was resuspended in 10 volumes/g lysisbuffer (50 mM HEPES, pH 7.2, containing 10% glycerol (v/v), 100 mM NaCl,2 mM DTT, 0.1 mM PMSF, 2 μg/ml Pepstatin, lug/ml each of E-64 andLeupeptin). Cells were lysed on ice using a microfluidizer andcentrifuged at 100,000×g for 30 min at 4° C. The 100,000×g supernatantwas diluted 1:5 with Buffer A (20 mM HEPES, pH 7.0, 10% glycerol (v/v),2 mM DTT) and purified by SP-Sepharose (Pharmacia) cation-exchangechromatography (column dimensions: 2.6×20 cm) at 4° C. The resin waswashed with 5 column volumes of Buffer A, followed by 5 column volumesof Buffer A containing 50 mM NaCl. Bound JNK3 was eluted with a 7.5column volume linear gradient of 50-300 mM NaCl. JNK3 eluted between150-200 mM NaCl.

Example 44 Activation of JNK3

Five mg of JNK3 was diluted to 0.5 mg/ml in 50 mM HEPES buffer, pH 7.5,containing 100 mM NaCl, 5 mM DTT, 20 mM MgCl₂ and 1 mM ATP. GST-MKK7(DD)was added at a molar ratio of 1:2.5 GST-MKK7:JNK3. After incubation for30 minutes at 25° C., the reaction mixture was concentrated 5-fold byultrafiltration in a Centriprep-30 (Amicon, Beverly, Mass.), diluted to10 ml and an additional 1 mM ATP added. This procedure was repeatedthree times to remove ADP and replenish ATP. The final addition of ATPwas 5 mM and the mixture incubated overnight at 4° C.

The activated JNK3/GST-MKK7(DD) reaction mixture was exchanged into 50mM HEPES buffer, pH 7.5, containing 5 mM DTT and 5% glycerol (w/v) bydialysis or ultrafiltration. The reaction mixture was adjusted to 1.1 Mpotassium phosphate, pH 7.5, and purified by hydrophobic interactionchromatography (at 25° C.) using a Rainin Hydropore column. GST-MKK7 andunactivated JNK3 do not bind under these conditions such that when a 1.1to 0.05 M potassium phosphate gradient is developed over 60 minutes at aflow rate of 1 ml/minute, doubly phosphorylated JNK3 is separated fromsingly phosphorylated JNK. Activated JNK3 (i.e. doubly phosphorylatedJNK3) was stored at −70° C. at 0.25-1 mg/ml.

Example 45 JNK Inhibition Assay

Compounds were assayed for the inhibition of JNK3 by aspectrophotometric coupled-enzyme assay. In this assay, a fixedconcentration of activated JNK3 (10 nM) was incubated with variousconcentrations of a potential inhibitor dissolved in DMSO for 10 minutesat 30° C. in a buffer containing 0.1 M HEPES buffer, pH 7.5, containing10 mM MgCl₂, 2.5 mM phosphoenolpyruvate, 200 μM NADH, 150 μg/mL pyruvatekinase, 50 μg/mL lactate dehydrogenase, and 200 μM EGF receptor peptide.The EGF receptor peptide has the sequence KRELVEPLTPSGEAPNQALLR, and isa phosphoryl acceptor in the JNK3-catalyzed kinase reaction. Thereaction was initiated by the addition of 10 μM ATP and the assay plateis inserted into the spectrophotometer's assay plate compartment thatwas maintained at 30° C. The decrease of absorbance at 340 nm wasmonitored as a function of time. The rate data as a function ofinhibitor concentration was fitted to competitive inhibition kineticmodel to determine the K_(i).

Table 13 shows the results of the activity of selected compounds of thisinvention in the JNK inhibition assay. The compound numbers correspondto the compound numbers in Tables 1, 2, and 7. Compounds having a K_(i)less than 0.1 micromolar (μM) are rated “A”, compounds having a K_(i)between 0.1 and 1 μM are rated “B” and compounds having a K_(i) greaterthan 1 μM are rated “C”. Compounds having an activity designated as “D”provided a percent inhibition less than or equal to 24%; compoundshaving an activity designated as “E” provided a percent inhibition ofbetween 24% and 66%; and compounds having an activity designated as “F”provided a provided a percent inhibition of between 67% and 100%.

TABLE 13 JNK3 Activity of Selected Compounds No. Activity No. ActivityNo. Activity IIa-1 D IIa-2 F IIa-3 D IIa-4 F IIa-5 D IIa-6 E IIa-7 DIIa-8 D IIa-9 D  IIa-10 A  IIa-11 A  IIa-12 E  IIa-13 B  IIa-14 A IIa-15 A  IIa-16 C — — — — IIb-1 D IIb-2 D IIb-3 D IIb-4 D IIb-5 DIIb-6 D  V-1 B — — — —

Example 46

The compounds were evaluated as inhibitors of human Src kinase usingeither a radioactivity-based assay or spectrophotometric assay.

Src Inhibition Assay A: Radioactivity-based Assay

The compounds were assayed as inhibitors of full length recombinanthuman Src kinase (from Upstate Biotechnology, cat. no. 14-117) expressedand purified from baculo viral cells. Src kinase activity was monitoredby following the incorporation of ³³p from ATP into the tyrosine of arandom poly Glu-Tyr polymer substrate of composition, Glu:Tyr=4:1(Sigma, cat. no. P-0275). The following were the final concentrations ofthe assay components: 0.05 M HEPES, pH 7.6, 10 mM MgCl₂, 2 mM DTT, 0.25mg/ml BSA, 10 μM ATP (1-2 uCi ³³P-ATP per reaction), 5 mg/ml polyGlu-Tyr, and 1-2 units of recombinant human Src kinase. In a typicalassay, all the reaction components with the exception of ATP werepre-mixed and aliquoted into assay plate wells.

Inhibitors dissolved in DMSO were added to the wells to give a finalDMSO concentration of 2.5%. The assay plate was incubated at 30° C. for10 min before initiating the reaction with ³³P-ATP. After 20 min ofreaction, the reactions were quenched with 150 μl of 10% trichloroaceticacid (TCA) containing 20 mM Na₃PO₄. The quenched samples were thentransferred to a 96-well filter plate (Whatman, UNI-Filter GF/F GlassFiber Filter, cat no. 7700-3310) installed on a filter plate vacuummanifold. Filter plates were washed four times with 10% TCA containing20 mM Na₃PO₄ and then 4 times with methanol. 200 μl of scintillationfluid was then added to each well. The plates were sealed and the amountof radioactivity associated with the filters was quantified on aTopCount scintillation counter. The radioactivity incorporated wasplotted as a function of the inhibitor concentration. The data wasfitted to a competitive inhibition kinetics model to get the K_(i) forthe compound.

Src Inhibition Assay B: Spectrophotometric Assay

The ADP produced from ATP by the human recombinant Src kinase-catalyzedphosphorylation of poly Glu-Tyr substrate was quanitified using acoupled enzyme assay (Fox et al (1998) Protein Sci 7, 2249). In thisassay one molecule of NADH is oxidised to NAD for every molecule of ADPproduced in the kinase reaction. The disappearance of NADH can beconveniently followed at 340 nm.

The following were the final concentrations of the assay components:0.025 M HEPES, pH 7.6, 10 mM MgCl2, 2 mM DTT, 0.25 mg/ml poly Glu-Tyr,and 25 nM of recombinant human Src kinase. Final concentrations of thecomponents of the coupled enzyme system were 2.5 mM phosphoenolpyruvate,200 μM NADH, 30 μg/ml pyruvate kinase and 10 μg/ml lactatedehydrogenase.

In a typical assay, all the reaction components with the exception ofATP were pre-mixed and aliquoted into assay plate wells. Inhibitorsdissolved in DMSO were added to the wells to give a final DMSOconcentration of 2.5%. The assay plate was incubated at 30 C. for 10 minbefore initiating the reaction with 100 μM ATP. The absorbance change at340 nm with time, the rate of the reaction, was monitored on a moleculardevices plate reader. The data of rate as a function of the inhibitorconcentration was fitted to compettive inhibition kinetics model to getthe K_(i) for the compound.

Table 14 shows the results of the activity of selected compounds of thisinvention in the Src inhibition assay. The compound numbers correspondto the compound numbers in Tables 1, 2, 7, and 8. Compounds having aK_(i) less than 0.1 micromolar (μM) are rated “A”, compounds having aK_(i) between 0.1 and 1 μM are rated “B” and compounds having a K_(i)greater than 1 μM are rated “C”. Compounds having an activity designatedas “D” provided a percent inhibition less than or equal to 24%;compounds having an activity designated as “E” provided a percentinhibition of between 24% and 66%; and compounds having an activitydesignated as “F” provided a provided a percent inhibition of between67% and 100%.

TABLE 14 Src Activity of Selected Compounds No. Activity No. ActivityNo. Activity IIa-1  D IIa-2  D IIa-3  D IIa-4  E IIa-5  D IIa-6  DIIa-7  D IIa-8  E IIa-9  D IIa-10 F IIa-11 F IIa-12 F IIa-13 F IIa-14 FIIa-15 F IIa-42 A IIa-43 A IIa-44 A IIa-49 B IIa-50 A IIa-68 A IIa-69 AIIa-71 A IIa-72 B IIa-73 B IIa-81 A IIa-82 B IIa-83 A IIa-86 B IIa-87 BIIa-88 B IIa-89 A IIa-90 C IIb-1  D IIb-2  D IIb-3  D IIb-4  E IIb-5  DIIb-6  D  V-4 A  V-5 A  V-15 A  V-29 A  V-30 A — —  VI-18 A  VI-19 A VI-20 A  VI-21 B  VI-22 A  VI-23 A  VI-24 A  VI-25 A  VI-117 B  VI-118B  VI-119 B  VI-120 A  VI-130 A  VI-132 B  VI-133 A  VI-134 A  VI-135 A VI-141 A

Example 47

The compounds were evaluated as inhibitors of human Lck kinase usingeither a radioactivity-based assay or spectrophotometric assay.

Lck Inhibition Assay A: Radioactivity-based Assay

The compounds were assayed as inhibitors of full length bovine thymusLck kinase (from Upstate Biotechnology, cat. no. 14-106) expressed andpurified from baculo viral cells. Lck kinase activity was monitored byfollowing the incorporation of ³³p from ATP into the tyrosine of arandom poly Glu-Tyr polymer substrate of composition, Glu:Tyr=4:1(Sigma, cat. no. P-0275). The following were the final concentrations ofthe assay components: 0.025 M HEPES, pH 7.6, 10 mM MgCl₂, 2 mM DTT, 0.25mg/ml BSA, 10 μM ATP (1-2 μCi ³³P-ATP per reaction), 5 mg/ml polyGlu-Tyr, and 1-2 units of recombinant human Src kinase. In a typicalassay, all the reaction components with the exception of ATP werepre-mixed and aliquoted into assay plate wells. Inhibitors dissolved inDMSO were added to the wells to give a final DMSO concentration of 2.5%.The assay plate was incubated at 30 C. for 10 min before initiating thereaction with ³³P-ATP. After 20 min of reaction, the reactions werequenched with 150 μl of 10% trichloroacetic acid (TCA) containing 20 mMNa₃PO₄. The quenched samples were then transferred to a 96-well filterplate (Whatman, UNI-Filter GF/F Glass Fiber Filter, cat no. 7700-3310)installed on a filter plate vacuum manifold. Filter plates were washedfour times with 10% TCA containing 20 mM Na₃PO₄ and then 4 times withmethanol. 200 μl of scintillation fluid was then added to each well. Theplates were sealed and the amount of radioactivity associated with thefilters was quantified on a TopCount scintillation counter. Theradioactivity incorporated was plotted as a function of the inhibitorconcentration. The data was fitted to a competitive inhibition kineticsmodel to get the K_(i) for the compound.

Lck Inhibition Assay B: Spectrophotometric Assay

The ADP produced from ATP by the human recombinant Lck kinase-catalyzedphosphorylation of poly Glu-Tyr substrate was quanitified using acoupled enzyme assay (Fox et al (1998) Protein Sci 7, 2249). In thisassay one molecule of NADH is oxidised to NAD for every molecule of ADPproduced in the kinase reaction. The disappearance of NADH can beconveniently followed at 340 nm.

The following were the final concentrations of the assay components:0.025 M HEPES, pH 7.6, 10 mM MgCl₂, 2 mM DTT, 5 mg/ml poly Glu-Tyr, and50 nM of recombinant human Lck kinase. Final concentrations of thecomponents of the coupled enzyme system were 2.5 mM phosphoenolpyruvate,200 μM NADH, 30 μg/ml pyruvate kinase and 10 μg/ml lactatedehydrogenase.

In a typical assay, all the reaction components with the exception ofATP were pre-mixed and aliquoted into assay plate wells. Inhibitorsdissolved in DMSO were added to the wells to give a final DMSOconcentration of 2.5%. The assay plate was incubated at 30° C. for 10min before initiating the reaction with 150 μM ATP. The absorbancechange at 340 nm with time, the rate of the reaction, was monitored on amolecular devices plate reader. The data of rate as a function of theinhibitor concentration was fitted to competitive inhibition kineticsmodel to get the K_(i) for the compound.

Table 15 shows the results of the activity of selected compounds of thisinvention in the Lck inhibition assay. The compound numbers correspondto the compound numbers in Tables 1, 7, and 8. Compounds having a K_(i)less than 0.1 micromolar (μM) are rated “A”, compounds having a K_(i)between 0.1 and 1 μM are rated “B” and compounds having a K_(i) greaterthan 1 μM are rated “C”.

TABLE 15 Lck Activity of Selected Compounds No. Activity No. ActivityNo. Activity IIa-10 A IIa-11 B IIa-14 A IIa-15 A IIa-38 A IIa-39 AIIa-40 A IIa-41 C IIa-42 A IIa-43 A IIa-44 A IIa-45 A IIa-46 A IIa-47 BIIa-48 A IIa-49 B IIa-50 A IIa-51 A IIa-52 C IIa-53 C IIa-54 C IIa-55 CIIa-56 C IIa-57 A IIa-58 A IIa-59 A IIa-60 A IIa-61 A IIa-62 C IIa-63 AIIa-64 A IIa-65 B IIa-66 A IIa-67 A IIa-68 A IIa-69 A IIa-70 A IIa-71 AIIa-72 B IIa-73 B IIa-74 C IIa-75 A IIa-76 C IIa-77 B IIa-78 C IIa-79 AIIa-80 C IIa-81 A IIa-82 C IIa-83 B IIa-84 C IIa-85 — IIa-86 A IIa-87 AIIa-88 B IIa-89 A IIa-90 B IIa-91 A IIa-92 A IIa-97 A IIa-98 A IIa-99 A IIa-100 A  IIa-101 A  IIa-105 A  IIa-106 A  IIa-107 B  IIa-108 A — — V-1 A  V-2 A  V-3 A  V-4 A  V-5 A  V-6 A  V-7 A  V-8 A  V-9 A  V-10 A V-11 A  V-12 B  V-13 A  V-14 B  V-15 A  V-16 B  V-17 B  V-18 A  V-19 A V-20 A  V-21 A  V-22 C  V-23 C  V-24 A  V-25 A  V-26 A  V-27 A  V-28 A V-29 A  V-30 A  V-31 C  V-32 A — — VI-1 A VI-2 A VI-3 A VI-4 A VI-5 AVI-6 A VI-7 A VI-8 A VI-9 A  VI-10 A  VI-11 A  VI-12 A  VI-13 A  VI-14 A VI-15 A  VI-16 A  VI-17 A  VI-18 A  VI-19 A  VI-20 A  VI-21 A  VI-22 B VI-23 C  VI-24 A  VI-25 B  VI-117 B  VI-118 A  VI-119 A  VI-120 A VI-121 B  VI-122 A  VI-123 B  VI-124 B  VI-125 B  VI-126 B  VI-127 B VI-128 A  VI-129 A  VI-130 A  VI-131 A  VI-132 A  VI-133 A  VI-134 A VI-135 A  VI-136 A  VI-137 A  VI-138 A  VI-139 A  VI-140 A  VI-141 A ——

While we have described a number of embodiments of this invention, it isapparent that our basic examples may be altered to provide otherembodiments which utilize the compounds and methods of this invention.Therefore, it will be appreciated that the scope of this invention is tobe defined by the appended claims rather than by the specificembodiments which have been represented by way of example.

We claim:
 1. A compound of formula I:

or a pharmaceutically acceptable salt thereof, wherein: A is N and B isCH; R¹ and R² are each independently selected from halogen, CN, NO₂,N(R)₂, OR, SR, or (T)_(n)—R⁵; R³ is selected from a 3-6 memberedcarbocyclic or heterocyclic ring having one to two heteroatomsindependently selected from nitrogen, oxygen, or sulfur, phenyl, or a5-6 membered heteroaryl ring having one to three heteroatomsindependently selected from nitrogen, oxygen, or sulfur, wherein saidphenyl or heteroaryl ring is optionally substituted with one (T)_(n)—Arand one to two R⁷; each n is independently selected from zero or one; Tis a C₁-C₆ alkylidene chain, wherein one methylene unit of T isoptionally replaced by CO, CO₂, COCO, CONR, OCONR, NRNR, NRNRCO, NRCO,NRCO₂, NRCONR, SO₂, NRSO₂, SO₂NR, NRSO₂NR, O, S, or NR; each R isindependently selected from hydrogen or an optionally substituted C₁-C₆aliphatic group; or two R on the same nitrogen atom may be takentogether with the nitrogen to form a four to eight membered, saturatedor unsaturated heterocyclic ring containing one to three heteroatomsindependently selected from nitrogen, oxygen, or sulfur; R⁴ is(T)_(n)—R, (T)_(n)—Ar, or (T)_(n)—Ar¹; R^(a) is selected from R^(b),halogen, NO₂, OR^(b), SR^(b), or N(R^(b))₂; R^(b) is selected fromhydrogen or a C₁-C₄ aliphatic group optionally substituted with oxo, OH,SH, NH₂, halogen, NO₂, or CN; R⁵ is an optionally substituted C₁-C₆aliphatic or Ar; Ar is a 5-6 membered saturated, partially unsaturated,or aryl monocyclic ring having zero to three heteroatoms independentlyselected from nitrogen, sulfur, or oxygen, or an 8-10 memberedsaturated, partially unsaturated, or aryl bicyclic ring having zero tofour heteroatoms independently selected from nitrogen, sulfur, oroxygen, wherein Ar is optionally substituted with one to three R⁷; Ar¹is a 6-membered aryl ring having zero to two nitrogens, wherein saidring is substituted with one Z—R⁶ group and optionally substituted withone to three R⁷; Z is a C₁-C₆ alkylidene chain wherein up to twonon-adjacent methylene units of Z are optionally replaced by CO, C₂,COCO, CONR, OCONR, NRNR, NRNRCO, NRCO, NRC₂, NRCONR, SO, SO₂, NRSO₂,SO₂NR, NRSO₂NR, O, S, or NR; provided that said optionally replacedmethylene unit of Z is a methylene unit non-adjacent to R⁶; R⁶ isselected from Ar, R, halogen, NO₂, CN, OR, SR, N(R)₂, NRC(O)R,NRC(O)N(R)₂, NRC₂R, C(O)R, CO₂R, OC(O)R, C(O)N(R)₂, OC(O)N(R)₂, SOR,SO₂R, SO₂N(R)₂, NRSO₂R, NRSO₂N(R)₂, C(O)C(O)R, or C(O)CH₂C(O)R; and eachR⁷ is independently selected from R, halogen, NO₂, CN, OR, SR, N(R)₂,NRC(O)R, NRC(O)N(R)₂, NRCO₂R, C(O)R, CO₂R, C(O)N(R)₂, OC(O)N(R)₂, SOR,SO₂R, SO₂N(R)₂, NRSO₂R, NRSO₂N(R)₂, C(O)C(O)R, or C(O)CH₂C(O)R; or twoR⁷ on adjacent positions of Ar¹ may be taken together to form asaturated, partially unsaturated, or fully unsaturated five to sevenmembered ring containing zero to three heteroatoms selected from O, S,or N.
 2. The compound according to claim 1, wherein said compound hasthe formula IIa:

or a pharmaceutically acceptable salt thereof.
 3. The compound accordingto claim 2, wherein said compound has one or more features selected fromthe group consisting of: (a) R¹ is selected from N(R)₂, OR, SR, or(T)_(n)—R⁵; (b) T is a C₁₋₄ alkylidene chain, wherein one methylene unitof T is optionally replaced by S, O, N(R), or CO₂; (c) R² is CN, R,halogen, C₂R⁵, or N(R)₂; (d) R³ is a 5-6 membered ring selected fromcarbocyclic, phenyl, or a heterocyclyl or heteroaryl ring having one totwo heteroatoms independently selected from nitrogen, oxygen or sulfur,wherein R³ is optionally substituted with one (T)_(n)—Ar group and oneR⁷; and (e) R⁴ is hydrogen or Ar, wherein Ar is an optionallysubstituted 6 membered saturated, partially saturated, or aryl ringhaving zero to two heteroatoms independently selected from nitrogen,oxygen, or sulfur.
 4. The compound according to claim 3, wherein saidcompound has one or more features selected from the group consisting of:(a) R¹ is selected from SCH₂-4-phenol, SCH₃, OH, OEt, N(Me)₂, OMe,4-methylpiperidin-1-yl, NHEt, NHCH₂CH₂piperidin-1-yl, orNHCH₂CH₂morpholin-4-yl; (b) R² is CN or CO₂R⁵; (c) R³ is selected fromphenyl, pyridyl, pyrimidinyl, cyclohexyl, or furanyl, wherein R³ isoptionally substituted with phenyl, phenoxy, benzyl, benzyloxy, pyridyl,3-hydroxyphenyl, 2-hydroxyphenyl, 3-aminophenyl, N-BOC-pyrrolyl,4-chlorophenyl, 3-ethoxypyridyl, 2-methoxypyridyl,2,5-dimethylisoxazolyl, 3-ethoxyphenyl, 4-isopropylphenyl,4-F-3-Cl-phenyl, pyrrolyl, pyrimidinyl, chloro, bromo, fluoro,trifluoromethyl, OH, NH₂, methyl, methoxy or ethoxy; and (d) R⁴ isselected from hydrogen or a phenyl, benzyl, pyridyl, piperidinyl, orcyclohexyl ring, wherein said ring is optionally subsituted withbenzyloxy, phenoxy, SO₂NH₂, OH, NO₂, NH₂, OMe, Br, Cl, CO₂Me, NHSO₂Me,NHSO₂Et, NHCON(Me)₂, NHCON(Et)₂, NHCOpyrrolidin-1-yl, orNHCOmorpholin-4-yl.
 5. The compound according to claim 1, wherein saidcompound has the formula IIb:

or a pharmaceutically acceptable sat thereof.
 6. The compound accordingto claim 5, wherein said compound has one or more features selected fromthe group consisting of: (a) R¹ is selected from N(R)₂, OR, SR, or(T)_(n)—R⁵; (b) T is a C₁₋₄ alkylidene chain, wherein one methylene unitof T is optionally replaced by S, O, N(R), or C₂; (c) R² is CN, R⁷, Ar,halogen, or N(R⁶)₂; (d) R³ is a 5-6 membered ring selected fromcarbocyclic, phenyl, or a heterocyclyl or heteroaryl ring having one totwo heteroatoms independently selected from nitrogen, oxygen or sulfur,wherein R³ is optionally substituted with one (T)_(n)—Ar group and oneR⁷; and (e) R⁴ is hydrogen or Ar, wherein Ar is an optionallysubstituted 6 membered saturated, partially saturated, or aryl ringhaving zero to two heteroatoms independently selected from nitrogen,oxygen, or sulfur.
 7. The compound according to claim 6, wherein saidcompound has one or more features selected from the group consisting of:(a) R¹ is selected from SCH₂-4-phenol, SCH₃, OH, OEt, N(Me)₂, OMe,4-methylpiperidin-1-yl, NHEt, NHCH₂CH₂piperidin-1-yl, orNHCH₂CH₂morpholin-4-yl; (b) R² is CN or 4-(C₁₋₃ alkyl)-thiazol-2-yl; (c)R³ is selected from phenyl, pyridyl, pyrimidinyl, cyclohexyl, orfuranyl, wherein R³ is optionally substituted with phenyl, phenoxy,benzyl, benzyloxy, pyridyl, 3-hydroxyphenyl, 2-hydroxyphenyl,3-aminophenyl, N-BOC-pyrrolyl, 4-chlorophenyl, 3-ethoxypyridyl,2-methoxypyridyl, 2,5-dimethylisoxazolyl, 3-ethoxyphenyl,4-isopropylphenyl, 4-F-3-Cl-phenyl, pyrrolyl, pyrimidinyl, chloro,bromo, fluoro, trifluoromethyl, OH, NH₂, methyl, methoxy or ethoxy; and(d) R⁴ is selected from hydrogen or a phenyl, benzyl, pyridyl,piperidinyl, or cyclohexyl ring, wherein said ring is optionallysubsituted with benzyloxy, phenoxy, SO₂NH₂, OH, NO₂, NH₂, OMe, Br, Cl,CO₂Me, NHSO₂Me, NHSO₂Et, NHCON(Me)₂, NHCON(Et)₂, NHCOpyrrolidin-1-yl, orNHCOmorpholin-4-yl.
 8. The compound according to claim 1, wherein saidcompound has the formula V:

or a pharmaceutically acceptable salt thereof.
 9. The compound accordingto claim 8, wherein said compound has one or more of the followingfeatures: (a) R¹ is selected from N(R)₂, OR, SR, or (T)_(n)—R⁵; (b) T isa C₁₋₄ alkylidene chain, wherein one methylene unit of T is optionallyreplaced by S, O, N(R), or C₂; (c) R² is CN, R, halogen, CO₂R⁵, orN(R)₂; (d) R³ is a 5-6 membered ring selected from carbocyclic, phenyl,or a heterocyclyl or heteroaryl ring having one to two heteroatomsindependently selected from nitrogen, oxygen or sulfur, wherein R³ isoptionally substituted with one (T)_(n)—Ar group and one R⁷; (e) R⁴ ishydrogen or Ar, wherein Ar is an optionally substituted 6 memberedsaturated, partially saturated, or aryl ring having zero to twoheteroatoms independently selected from nitrogen, oxygen, or sulfur; and(f) R^(a) is selected from R^(b), OR^(b), SR^(b), or N(R^(b))₂.
 10. Thecompound according to claim 9, wherein said compound has one or more ofthe following features: (a) R¹ is selected from SCH₂-4-phenol, SCH₃, OH,OEt, N(Me)₂, OMe, 4-methylpiperidin-1-yl, NHEt, NHCH₂CH₂piperidin-1-yl,or NHCH₂CH₂morpholin-4-yl; (b) R² is CN or CO₂R⁵; (c) R³ is selectedfrom phenyl, pyridyl, pyrimidinyl, cyclohexyl, or furanyl, wherein R³ isoptionally substituted with phenyl, phenoxy, benzyl, benzyloxy, pyridyl,3-hydroxyphenyl, 2-hydroxyphenyl, 3-aminophenyl, N-BOC-pyrrolyl,4-chlorophenyl, 3-ethoxypyridyl, 2-methoxypyridyl,2,5-dimethylisoxazolyl, 3-ethoxyphenyl, 4-isopropylphenyl,4-F-3-Cl-phenyl, pyrrolyl, pyrimidinyl, chloro, bromo, fluoro,trifluoromethyl, OH, NH₂, methyl, methoxy or ethoxy; (d) R⁴ is selectedfrom hydrogen or a phenyl, benzyl, pyridyl, piperidinyl, or cyclohexylring, wherein said ring is optionally subsituted with benzyloxy,phenoxy, SO₂NH₂, OH, NO₂, NH₂, OMe, Br, Cl, CO₂Me, NHSO₂Me, NHSO₂Et,NHCON(Me)₂, NHCON(Et)₂, NHCOpyrrolidin-1-yl, or NHCOmorpholin-4-yl; and(e) R^(a) is methyl, OH, OMe, or NH₂.
 11. The compound according toclaim 1, wherein said compound has the formula VI:

or a pharmaceutically acceptable salt thereof.
 12. The compoundaccording to claim 11, wherein said compound has one or more featuresselected from the group consisting of: (a) R¹ is N(R)₂, OR, SR, or(T)_(n)—R⁵; (b) T is a C₁₋₄ alkylidene chain, wherein one methylene unitof T is optionally replaced by S, O, N(R), or CO₂; (c) R² is CN, R⁷,halogen, or N(R⁶)₂; (d) R³ is a 5-6 membered ring selected fromcarbocyclic, phenyl, or a heterocyclyl or heteroaryl ring having one totwo heteroatoms independently selected from nitrogen, oxygen or sulfur,wherein R³ is optionally substituted with one (T)_(n)—Ar group and oneR⁷; (e) Z is a C₁₋₄ alkylidene chain wherein one methylene unit of Z isoptionally replaced by O, NH, NHCO, NHCO₂, NHSO₂, CONH; (f) R⁶ isselected from N(R)₂, NHCOR, or Ar wherein Ar is an optionallysubstituted 5-6 membered heterocyclic or heteroaryl ring having one totwo heteroatoms independently selected from nitrogen, oxygen, or sulfur;and (g) R^(a) is R^(b), OR^(b), SR^(b), or N(R^(b))₂.
 13. The compoundaccording to claim 12, wherein said compound has one or more featuresselected from the group consisting of: (a) R¹ is selected fromSCH₂-4-phenol, SCH₃, OH, OEt, N(Me)₂, OMe, 4-methylpiperidin-1-yl, NHEt,NHCH₂CH₂piperidin-1-yl, or NHCH₂CH₂morpholin-4-yl; (b) R² is CN; (c) R³is a phenyl, pyridyl, furyl, or cyclohexyl ring optionally substitutedwith (T)_(n)—Ar or R⁷ wherein Ar is a 5-6 membered aryl ring having zeroto two heteroatoms independently selected from nitrogen, oxygen, orsulfur, and wherein R⁷ is selected from R, halogen, OR, N(R)₂, or CO₂R;(d) R^(a) is hydrogen or methyl; and (e) Z—R⁶ is selected fromO(CH₂)₃OH, O(CH₂)₃NH(CH₂)₂OH, O(CH₂)₂NH(CH₂)₂OH,O(CH₂)₃N(hydroxyethyl)(methyl), O(CH₂)₃pyrrolidin-1-yl,O(CH₂)₂morpholin-4-yl, O(CH₂)₃N(Me)₂, O(CH₂)₃N(Et)₂,O(CH₂)₃(4-hydroxyethylpiperazin-1-yl), O(CH₂)₃piperazin-1-yl,O(CH₂)₃(4-hydroxymethylpiperidin-1-yl),O(CH₂)₃(4-hydroxypiperidin-1-yl), NHCO(CH₂)₃N(Me)₂, NHCO(CH₂)₃NCOCH₃,NHCOCH₂pyridin-2-yl, NHCOCH₂(2-aminothiazol-4-yl), NHCOCH₂cyclopropyl,NHCO(CH₂)₂N(Et)₂, NHCO(CH₂)₂(piperazin-2,5-dione-3-yl),NHCOpyrrolidin-1-yl, NHCOmorpholin-4-yl, NHCO₂CH₂tetrahydrofuran-2-yl,NHCO₂tetrahydrofuran-2-yl, NHCO₂tetrahydropyran-4-yl, orNHCO₂CH₂tetrahydropyran-2-yl.
 14. A compound selected from the followingTable 1, Table 2, Table 7, or Table 8 compounds: TABLE 1 Compounds ofFormula IIa

No. IIa- R¹ R² R³ R⁴ 1 SMe CN 4-CO₂H-phenyl H 2 SMe CN 4-Cl-phenyl H 3SMe CN 4-CF₃-phenyl H 4 SMe CN 4-CH₃-phenyl H 5 SMe CN 2-Cl-phenyl H 6SMe CN 4-OCH₃-phenyl H 7 NHCH₂Ph CN 4-CF₃-phenyl H 8 NHCH₂Ph CN2-Cl-phenyl H 9 NHCH₂Ph CN 4-OCH₃-phenyl H 10 SMe CN 4-Cl-phenyl Ph 11OEt CN 4-Cl-phenyl Ph 12 SMe CN 4-CF₃-phenyl Ph 13 OEt CN 4-CF₃-phenylPh 14 SMe CN 4-CH₃-phenyl Ph 15 OEt CN 4-CH₃-phenyl Ph 16 SMe CN H H 17CH₂CH₂OH CN OPh Et 18 CONHEt CF₃ pyridine-3-yl CH₂Ph 19 SCH₂Ph NHEtCONHCH₂Ph COPh 20 CH₂NO₂ CONHEt NH(4-Cl-phenyl) H 21 NHCONH₂ OMe CH₂PhSO₂Me 22 Et CN thiazol-2-yl Ph 23 SMe CN piperidin-1-yl cyclohexyl 24OCH₂Ph Cl 4-CONHMe-phenyl cyclohexyl 25 NHMe NO₂ NHPh H 26 SMe NO₂ NH₂ H27 OEt NO₂ NHCH₂Ph H 28 NHMe NH₂ NHPh Ph 29 SMe NH₂ NH₂ Me 30 OEt NH₂NHCH₂Ph Me 31 NHMe NHCOEt Ph Ph 32 SME NHCOEt CH₂Ph CH₂Ph 33 OEt NHCOEtCH₂Ph H 34 NHMe CONHMe Ph H 35 SMe CONHMe Ph Ph 36 OEt CONHMe 4-Cl—Ph Ph37 SMe CN Ph 3-OBn—Ph 38 SMe CN Ph 3-SO₂NH₂—Ph 39 S—CH₂- CN Ph Phphenol-4-ol 40 SMe CN Ph 3-OH—Ph 41 SMe CN Ph 4-OBn—Ph 42 OH CN Ph3-OBn—Ph 43 SMe CN cyclohexyl 3,5-OMe—Ph 44 SMe CN cyclohexyl3-SO₂NH₂—Ph 45 SMe CN cyclohexyl 3-OBn—Ph 46 SMe CN cyclohexyl Ph 47 SMeCN cyclohexyl 4-CO₂Et—Ph 48 SMe CN cyclohexyl 3-OH—Ph 49 SMe CN 3-OMe—Ph3-NO₂—Ph 50 SMe CN 3-OMe—Ph 3-NH₂—Ph 51 SMe CN 3-OH—Ph 3-NO₂—Ph 52 SMeCN 3-OBn—Ph Ph 53 SMe CN 3-OBn—Ph 3-NO₂—Ph 54 N(Me)₂ CN 3-OBn—Ph Ph 55N(Me)₂ CN 3-OBn—Ph 3-NO₂—Ph 56 SMe CN 3-pyridyl 3-OBn—Ph 57 SMe CN3-pyridyl 3-OH—Ph 58 OEt CN Ph Ph 59 SMe CN 3-Br—Ph 3-NH₂—Ph 60 N(Me)₂CN 3-OPh—Ph 3-NH₂—Ph 61 SMe CN 3-OPh—Ph 3-NH₂—Ph 62 SMe CN5-Br-3-pyridyl 3-OBn—Ph 63 4-Me-piper- CN 3-OPh—Ph Ph idin-1-yl 64 OH CN4-tolyl Ph 65 SMe CN 3-OBn—Ph 3-OH—Ph 66 SMe CN 3-OPh—Ph 3-OH—Ph 67 SMeCN 3-OH—Ph 3-OH—Ph 68 SMe CN 3-Br—Ph 3-OH—Ph 69 SMe CN 3-Br—Ph 3-OBn—Ph70 NHEt CN Ph 3-OH—Ph 71 SMe CN 3-(3-OH—Ph)—Ph 3-OH—Ph 72 SMe CN3-(3-OEt—Ph)—Ph 3-OH—Ph 73 SMe CN 3-(3-pyridyl)-Ph 3-OH—Ph 74 SMe CN5-Ph-pyridin-3-yl 3-OBn—Ph 75 N(Me)₂ CN 5-Br-3-pyridyl 3-OBn—Ph 76N(Me)₂ CN 5-Ph-3-pyridyl 3-OBn—Ph 77 SMe CO₂Et Ph 3-OH—Ph 78 SMe CO₂EtPh Ph 79 NHEt CN Ph 4-OH—Ph 80 N(Me)₂ CN 5-Ph-pyridin-3-yl 3-OH—Ph 81SMe CN 3-(3-NH₂—Ph)—Ph 3-OH—Ph 82 SMe CN 3-(3-Cl,4-F—Ph)—Ph 3-OH—Ph 83SMe CN 3-(4-iPr—Ph)—Ph 3-OH—Ph 84 SMe CN 5-Ph-pyridin-3-yl 3-OH—Ph 85SMe CN 5-Ph-pyridin-3-yl 3-NO₂—Ph 86 SMe CN 3-(3-N—Boc-pyrrol-2- 3-OH—Phyl)-Ph 87 SMe CN 3-(4-iPr—Ph)—Ph 3-OH—Ph 88 SMe CN 3-pyridyl3-NHSO₂Me—Ph 89 SMe CN 3-pyridyl 3-NHSO₂Et—Ph 90 NHEt CO₂Et Ph 3-OMe—Ph91 SMe CN 3-pyridyl 3-SO₂NH₂—Ph 92 SMe CN 3-(2-OH—Ph)—Ph 3-OH—Ph 93 SMeCN 3-(3-pyrrol-2-yl)-Ph 3-OH—Ph 94 SMe CN 3-(6-OMe-pyridin-2- 3-OH—Phyl)-Ph 95 SMe CN 3-(5-OMe-pyridin-2- 3-OH—Ph yl)-Ph 96 SMe CN3-(2,5-Me₂-isoxazol-4- 3-OH—Ph yl)-Ph 97 NH(CH₃)₂mor- CN Ph 3-OH—Phpholin-4-yl 98 NH(CH₃)₂mor- CN Ph 3-SO₂NH₂—Ph pholin-4-yl 99NH(CH₃)₂mor- CN Ph 4-OH—Ph pholin-4-yl 100 NH(CH₃)₃N(Et)₂ CN Ph 3-OH—Ph101 NH(CH₃)₃N(Et)₂ CN Ph 4-OH—Ph 102 NH(CH₃)₂piperidin- CN Ph 3-OH—Ph1-yl 103 NH(CH₃)₂piperidin- CN Ph 3-SO₂NH₂—Ph 1-yl 104NH(CH₃)₂piperidin- CN Ph 4-OH—Ph 1-yl 105 SMe CN 3-(pyridin-4-yl)-Ph3-OH—Ph

TABLE 2 Compounds of Formula IIb

No. IIb- R¹ R² R³ R⁴ 1 SMe 4-CH₃thiazol-2-yl Me Ph 2 SMe4-CH₃thiazol-2-yl Me 4-F—Ph 3 SMe 4-CH₃thiazol-2-yl Me 6-Cl-pyridin-3-yl4 SMe 4-CH₃thiazol-2-yl Me 3-Cl—Ph 5 SMe 4-CH₃thiazol-2-yl Me 3-CH₃O—Ph6 SMe 4-CH₃thiazol-2-yl Me 3-BnO—Ph 7 SMe CN Me Ph 8 SMe CN Me 4-F—Ph 9SMe CN Me 6-Cl-pyridin-3-yl 10 SMe CN Me 3-Cl—Ph 11 SMe CN Me 3-CH₃O—Ph12 SMe CN Me 3-BnO—Ph

TABLE 7 Compounds of Formula V

No.V- R¹ R^(a) R³ R⁴ 1 SCH₂phen-4-ol Me 3-Cl phenyl Ph 2 OH Me Ph3-OBn—Ph 3 OEt Me Ph 3-OBn—Ph 4 SMe Me Ph 3-OBn—Ph 5 SMe Me Ph3-SO₂NH₂—Ph 6 SMe Me Ph 3-OH—Ph 7 SMe Me Ph 3-NO₂—Ph 8 SMe Me Ph3-NH₂—Ph 9 SMe Me 2-CF₃—Ph 3-OBn—Ph 10 SMe Me 2-CF₃—Ph 3-OH—Ph 11SCH₂phen-4-ol Me 3-Cl—Ph Ph 12 SMe Me 4-Me—Ph 3-OBn—Ph 13 SMe Me 4-Me—Ph3-OH—Ph 14 SMe Me 4-Me—Ph pyrid-3-yl 15 SMe Me pyrid-3-yl 3-OBn—Ph 16SMe Me pyrid-3-yl Ph 17 SMe Me pyrid-3-yl 3-OMe—Ph 18 SMe Me pyrid-3-yl3,5-OMe—Ph 19 SMe Me pyrid-3-yl 3-Br—Ph 20 SMe Me pyrid-3-yl 3-Cl—Ph 21SMe Me pyrid-3-yl 3-CO₂Me—Ph 22 SMe Me pyrid-3-yl 6-Cl-pyrid-3-yl 23 SMeMe pyrid-3-yl CH₂Ph 24 SMe Me pyrid-3-yl 3-OH—Ph 25 SMe Me furan-2-ylpyrid-3-yl 26 SMe Me furan-2-yl 3-OH—Ph 27 SMe Me furan-2-yl 3-OBn—Ph 28SMe Me furan-2-yl 3-NO₂—Ph 29 N(Me)₂ Me 3-OPh—Ph 3-NO₂—Ph 30 N(Me)₂ Me3-OPh—Ph 3-OH—Ph 31 SMe Me 3-Ph—Ph 3-OBn—Ph 32 SMe Me 3-Ph—Ph 3-OH—Ph

TABLE 8


15. A composition comprising an effective amount of a compound accordingto claim 1, and a pharmaceutically acceptable carrier, adjuvant, orvehicle.
 16. The composition according to claim 15, additionallycomprising a therapeutic agent selected from an anti-proliferativeagent, an anti-inflammatory agent, an immunomodulatory agent, an agentfor treating neurological disorders, an agent for treatingcardiovascular disease, an agent for treating liver disease, ananti-viral agent, an agent for treating blood disorders, an agent fortreating diabetes, or an agent for treating immunodeficiency disorders.17. A composition for coating an implantable device comprising acompound according to claim 1 and a carrier suitable for coating saidimplantable device.
 18. A method of inhibiting JNK, Lck, or Src kinaseactivity in a biological sample comprising the step of contacting saidbiological sample with: a) a compound according to claim 1; or b) acomposition according to claim
 15. 19. A method of treating or lesseningthe severity of an inflammatory disease, autoimmune disease, destructivebone disorder, proliferative disorder, infectious disease,neurodegenerative disease, allergy, reperfusion/ischemia in stroke,heart attack, angiogenic disorder, organ hypoxia, vascular hyperplasia,cardiac hypertrophy, thrombin-induced platelet aggregation or acondition associated with proinflammatory cytokines, comprising the stepof administering to said patient a composition according to claim 15.20. The method according to claim 19, wherein said method is used totreat or lessen the severity of an inflammatory disease selected fromacute pancreatitis, chronic pancreatitis, asthma, allergies, or adultrespiratory distress syndrome.
 21. The method according to claim 20,wherein said method is used to treat or lessen the severity of anautoimmune disease selected from glomerulonephritis, rheumatoidarthritis, systemic lupus erythematosus, scleroderma, chronicthyroiditis, Graves' disease, autoimmune gastritis, diabetes, autoimmunehemolytic anemia, autoimmune neutropenia, thrombocytopenia, atopicdermatitis, chronic active hepatitis, myasthenia gravis, multiplesclerosis, inflammatory bowel disease, ulcerative colitis, Crohn'sdisease, psoriasis, or graft vs. host disease.
 22. The method accordingto claim 20, wherein said method is used to treat or lessen the severityof a destructive bone disorders selected from osteoarthritis,osteoporosis or multiple myeloma-related bone disorder.
 23. The methodaccording to claim 20, wherein said method is used to treat or lessenthe severity of a proliferative disease selected from acute myelogenousleukemia, chronic myelogenous leukemia, metastatic melanoma, Kaposi'ssarcoma, or multiple myeloma.
 24. The method according to claim 20,wherein said method is used to treat or lessen the severity of aneurodegenerative disease selected from Alzheimer's disease, Parkinson'sdisease, amyotrophic lateral sclerosis, Huntington's disease, cerebralischemia or neurodegenerative disease caused by traumatic injury,glutamate neurotoxicity or hypoxia.
 25. The method according to claim20, wherein said method is used to treat or lessen the severity ofischemia/reperfusion in stroke or myocardial ischemia, renal ischemia,heart attacks, organ hypoxia or thrombin-induced platelet aggregation.26. The method according to claim 20, wherein said method is used totreat or lessen the severity of a condition associated with T-Cellactivation or pathologic immune responses.
 27. The method according toclaim 20, wherein said method is used to treat or prevent an angiogenicdisorder selected from solid tumors, ocular neovasculization, orinfantile haemangiomas.
 28. The method according to claim 20, whereinsaid disease is selected from hypercalcemia, restenosis, hypercalcemia,osteoporosis, osteoarthritis, symptomatic treatment of bone metastasis,rheumatoid arthritis, inflammatory bowel disease, multiple sclerosis,psoriasis, lupus, graft vs. host disease, T-Cell mediatedhypersensitivity disease, Hashimoto's thyroiditis, Guillain-Barresyndrome, chronic obtructive pulmonary disorder, contact dermatitis,cancer, Paget's disease, asthma, ischemic or reperfusion injury,allergic disease, atopic dermatitis, or allergic rhinitis.
 29. Themethod according to claim 28, wherein said disease is selected fromhypercalcemia, osteoperosis, osteoarthritis, or sympomatic treatment ofbone metastasis.
 30. The method according to claim 19, wherein saiddisease is selected from autoimmune diseases, allergies, rheumatoidarthritis, and leukemia.
 31. The method according to claim 19,comprising the additional step of administering to said patient anadditional therapeutic agent selected from an anti-proliferative agent,an anti-inflammatory agent, an immunomodulatory agent, an agent fortreating neurological disorders, an agent for treating cardiovasculardisease, an agent for treating liver disease, an anti-viral agent, anagent for treating blood disorders, an agent for treating diabetes, oran agent for treating immunodeficiency disorders, wherein: saidadditional therapeutic agent is appropriate for the disease beingtreated; and said additional therapeutic agent is administered togetherwith said composition as a single dosage form or separately from saidcomposition as part of a multiple dosage form.